B , C HeLa cells stably expressing GFP - Nup133 were treated with the indicated siRNAs , synchronized by double thymidine block , released for 12 hours , treated with or without 1 , 6 Hexanediol and analysed by immunofluorescence microscopy . The magnified framed regions are shown in the corresponding numbered panels . The percentage of cells with cytoplasmic GFP - Nup133 granules was quantified in ( C ) , 3100 cells were analysed ( mean ± SD , * * P < 0 . 01 ; * * * P < 0 . 001 ; * * * * P < 0 . 0001 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B, C HeLa cells stably expressing GFP-Nup133 were treated with the indicated siRNAs, synchronized by double thymidine block, released for 12 hours, treated with or without 1,6 Hexanediol and analysed by immunofluorescence microscopy. The magnified framed regions are shown in the corresponding numbered panels. The percentage of cells with cytoplasmic GFP-Nup133 granules was quantified in (C), 3100 cells were analysed (mean ±SD, **P < 0.01; ***P < 0.001; ****P < 0.0001; N = 3).


A HeLa cells stably expressing GFP alone or GFP - FXR1 were immunoprecipitated using GFP - Trap beads ( GFP - IP ) , analysed by Western blot and quantified ( mean , * P < 0 . 05 ; N = 3 ) .

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A HeLa cells stably expressing GFP alone or GFP-FXR1 were immunoprecipitated using GFP-Trap beads (GFP-IP), analysed by Western blot and quantified (mean, *P < 0.05; N = 3).


B , C HeLa cells were treated with the indicated siRNAs , synchronized by double thymidine block , released for 12 hours and analysed by immunofluorescence microscopy . Arrowheads indicate blebbed regions of nuclei . The percentage of interphasic cells with Nup granules was quantified ( C ) , 900 cells were analysed ( mean ± SD , * * * P < 0 . 001 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B, C HeLa cells were treated with the indicated siRNAs, synchronized by double thymidine block, released for 12 hours and analysed by immunofluorescence microscopy. Arrowheads indicate blebbed regions of nuclei. The percentage of interphasic cells with Nup granules was quantified (C), 900 cells were analysed (mean ±SD, ***P < 0.001; N = 3).


D Lysates of HeLa cells stably expressing GFP alone or GFP - FXR1 were subjected to immunoprecipitation using GFP - Trap beads ( GFP - IP ) , analysed by Western blot and quantified ( mean , * P < 0 . 05 ; * * P < 0 . 01 ; N = 3 ) .

O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

D Lysates of HeLa cells stably expressing GFP alone or GFP-FXR1 were subjected to immunoprecipitation using GFP-Trap beads (GFP-IP), analysed by Western blot and quantified (mean, *P < 0.05; **P < 0.01; N = 3).


E , F HeLa cells were treated with the indicated siRNAs , synchronized by double thymidine block and released for 12 hours and analysed by immunofluorescence microscopy for FXR1 and mAb414 . The magnified framed regions are shown in the corresponding numbered panels in ( E ) . The percentage of cells with cytoplasmic Nup granules was quantified in ( F ) , 2700 cells were analysed ( mean ± SD , * * * P < 0 . 001 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

E, F HeLa cells were treated with the indicated siRNAs, synchronized by double thymidine block and released for 12 hours and analysed by immunofluorescence microscopy for FXR1 and mAb414. The magnified framed regions are shown in the corresponding numbered panels in (E). The percentage of cells with cytoplasmic Nup granules was quantified in (F), 2700 cells were analysed (mean ±SD, ***P < 0.001; N = 3).


A , B HeLa cells stably expressing GFP - Nup133 were treated with the indicated siRNAs , synchronized by double thymidine block , released for 12 hours , treated with nocodazole to induce granule formation and washed - out as indicated and analysed by immunofluorescence microscopy . The percentage of cells with cytoplasmic GFP - Nup133 granules was quantified in ( B ) , 5200 cells were analysed ( mean ± SD , * * P < 0 . 01 ; * * * P < 0 . 001 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

A, B HeLa cells stably expressing GFP-Nup133 were treated with the indicated siRNAs, synchronized by double thymidine block, released for 12 hours, treated with nocodazole to induce granule formation and washed-out as indicated and analysed by immunofluorescence microscopy. The percentage of cells with cytoplasmic GFP-Nup133 granules was quantified in (B), 5200 cells were analysed (mean ±SD, **P < 0.01; ***P < 0.001; N = 3).


C - E HeLa cells stably expressing GFP - Nup107 were treated with the indicated siRNAs , synchronized by double thymidine block , released for 12 hours , treated with nocodazole to induce granule formation and washed - out as in ( A ) and analysed by live video spinning disk confocal microscopy . The selected frames of the movies are depicted and time is shown in minutes . The magnified framed regions are depicted in the lower rows . White arrowheads point to individual GFP - Nup107 - positive granules . Yellow arrowheads point to the granules undergoing fusion events . The percentage of cells with fusion / fission events of GFP - Nup107 granules was quantified in ( D ) , 815 cells were analysed ( mean ± SD , * * P < 0 . 01 ; N = 3 ) . The number of fusion / fission events per cell was quantified in ( E ) , 815 cells were analysed ( mean ± SD ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

C-E HeLa cells stably expressing GFP-Nup107 were treated with the indicated siRNAs, synchronized by double thymidine block, released for 12 hours, treated with nocodazole to induce granule formation and washed-out as in (A) and analysed by live video spinning disk confocal microscopy. The selected frames of the movies are depicted and time is shown in minutes. The magnified framed regions are depicted in the lower rows. White arrowheads point to individual GFP-Nup107-positive granules. Yellow arrowheads point to the granules undergoing fusion events. The percentage of cells with fusion/fission events of GFP-Nup107 granules was quantified in (D), 815 cells were analysed (mean ±SD, **P < 0.01; N = 3). The number of fusion/fission events per cell was quantified in (E), 815 cells were analysed (mean ±SD; N = 3).


Human FXS - patient derived fibroblasts ( FXS - 1 , FXS - 2 ) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by Western blot ( A )

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Human FXS-patient derived fibroblasts (FXS-1, FXS-2) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by Western blot (A)


Human FXS - patient derived fibroblasts ( FXS - 1 , FXS - 2 ) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy Examples of Nup localization defects are shown in ( B )

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O

Human FXS-patient derived fibroblasts (FXS-1, FXS-2) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy Examples of Nup localization defects are shown in (B)


Human FXS - patient derived fibroblasts ( FXS - 1 , FXS - 2 ) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy The percentage of cells with cytoplasmic Nup granules was quantified in ( C ) , 283 cells were analysed ( mean ± SD , * * * P < 0 . 001 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Human FXS-patient derived fibroblasts (FXS-1, FXS-2) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy The percentage of cells with cytoplasmic Nup granules was quantified in (C), 283 cells were analysed (mean ±SD, ***P < 0.001; N = 3).


Human FXS - patient derived fibroblasts ( FXS - 1 , FXS - 2 ) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy examples of nuclear architecture defects are shown in ( D ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Human FXS-patient derived fibroblasts (FXS-1, FXS-2) and control human fibroblasts were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy examples of nuclear architecture defects are shown in (D).


Human induced pluripotent stem cells ( iPSCs ) derived from a FXS patient ( FXS - iPSC ) and the isogenic rescue cells ( C1 _ 2 - iPSC ) were analysed by immunofluorescence

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Human induced pluripotent stem cells (iPSCs) derived from a FXS patient (FXS-iPSC) and the isogenic rescue cells (C1_2-iPSC) were analysed by immunofluorescence microscopy


Human induced pluripotent stem cells ( iPSCs ) derived from a FXS patient ( FXS - iPSC ) and the isogenic rescue cells ( C1 _ 2 - iPSC ) were analysed by immunofluorescence microscopy The percentage of cells with cytoplasmic Nup133 granules was quantified in ( F ) , 5500 cells were analysed ( mean ± SD , * * P < 0 . 01 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Human induced pluripotent stem cells (iPSCs) derived from a FXS patient (FXS-iPSC) and the isogenic rescue cells (C1_2-iPSC) were analysed by immunofluorescence microscopy The percentage of cells with cytoplasmic Nup133 granules was quantified in (F), 5500 cells were analysed (mean ±SD, **P < 0.01; N = 3).


Human induced pluripotent stem cells ( iPSCs ) derived from a FXS patient ( FXS - iPSC ) and the isogenic rescue cells ( C1 _ 2 - iPSC ) were analysed by immunofluorescence microscopy Examples of co - localization events of re - expressed FMRP and Nup133 are shown in ( G ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O

Human induced pluripotent stem cells (iPSCs) derived from a FXS patient (FXS-iPSC) and the isogenic rescue cells (C1_2-iPSC) were analysed by immunofluorescence microscopy Examples of co-localization events of re-expressed FMRP and Nup133 are shown in (G).


H , I Mouse Embryonic Fibroblasts ( MEFs ) derived from the Fmr1 knock - out ( KO ) mice and wild type controls were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy ( H ) . The percentage of cells with cytoplasmic nucleoporin granules was quantified in ( I ) , 2400 cells were analysed ( mean ± SD , * P < 0 . 05 ; N = 3 ) .

O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

H, I Mouse Embryonic Fibroblasts (MEFs) derived from the Fmr1 knock-out (KO) mice and wild type controls were synchronized in early G1 by Monastrol release and analysed by immunofluorescence microscopy (H). The percentage of cells with cytoplasmic nucleoporin granules was quantified in (I), 2400 cells were analysed (mean ±SD, *P < 0.05; N = 3).


A HeLa cells were transfected with the import / export reporter plasmid XRGG - GFP , treated with the indicated siRNAs and synchronized in early G1 phase by Monastrol release . Dexamethasone was added for 3 hours to induce XRGG - GFP nuclear import . Following wash - out the nuclear export of XRGG - GFP was analysed by live video spinning disk confocal microscopy . The selected frames of the movies are depicted in Figure EV5C . The percentage of cytoplasmic XRGG - GFP over time was quantified in Figure EV5D and quantifications of individual cells from the 20 and 30 minutes time points are depicted in ( A ) , 199 cells were analysed ( mean ± SD , * P < 0 . 05 ; * * P < 0 . 01 ; N = 3 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

A HeLa cells were transfected with the import/export reporter plasmid XRGG-GFP, treated with the indicated siRNAs and synchronized in early G1 phase by Monastrol release. Dexamethasone was added for 3 hours to induce XRGG-GFP nuclear import. Following wash-out the nuclear export of XRGG-GFP was analysed by live video spinning disk confocal microscopy. The selected frames of the movies are depicted in Figure EV5C. The percentage of cytoplasmic XRGG-GFP over time was quantified in Figure EV5D and quantifications of individual cells from the 20 and 30 minutes time points are depicted in (A), 199 cells were analysed (mean ±SD, *P < 0.05; **P < 0.01; N = 3).


B HeLa cells stably expressing GFP - Nup133 were treated with the indicated siRNAs , synchronized by double thymidine block , released for 12 hours and analysed by immunofluorescence microscopy . The magnified framed regions are shown in the corresponding numbered panels .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B HeLa cells stably expressing GFP-Nup133 were treated with the indicated siRNAs, synchronized by double thymidine block, released for 12 hours and analysed by immunofluorescence microscopy. The magnified framed regions are shown in the corresponding numbered panels.


C , D Asynchronously proliferating HeLa cells were treated with indicated siRNAs and analysed by immunofluorescence microscopy ( C ) . The percentage of p - Rb positive cells was quantified in ( D ) , 2800 cells were analysed ( mean ± SD , * * P < 0 . 01 ; N = 3 ) .

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C, D Asynchronously proliferating HeLa cells were treated with indicated siRNAs and analysed by immunofluorescence microscopy (C). The percentage of p-Rb positive cells was quantified in (D), 2800 cells were analysed (mean ±SD, **P < 0.01; N = 3).


Asynchronously proliferating HeLa cells were treated with indicated siRNAs , incubated with EdU during 30min and analysed by immunofluorescence microscopy . The percentage of p - Rb and / or EdU positive cells was quantified in ( E ) , 2100 cells were analysed ( mean ± SD , * P < 0 . 05 ; * * P < 0 . 01 ; N = 3 )

O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Asynchronously proliferating HeLa cells were treated with indicated siRNAs, incubated with EdU during 30min and analysed by immunofluorescence microscopy. The percentage of p-Rb and/or EdU positive cells was quantified in (E), 2100 cells were analysed (mean ±SD, *P < 0.05; **P < 0.01; N = 3)


F Asynchronously proliferating HeLa cells were treated with indicated siRNAs , incubated with EdU during 30min and analysed by immunofluorescence microscopy . the percentage of p - Rb and / or cyclin B positive cells was quantified in (

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F Asynchronously proliferating HeLa cells were treated with indicated siRNAs, incubated with EdU during 30min and analysed by immunofluorescence microscopy. the percentage of p-Rb and/or cyclin B positive cells was quantified in (


A B16F10 melanoma cells with either mTRF2 overexpression or knockdown were inoculated into the backs of immunocompetent mice . The percentages of intratumoral CD11bHi GR1Hi MDSCs ( n = 10 mice ) , CD25 + Foxp3 + regulatory T cells ( Tregs ) , or CD25 + Foxp3 − activated T cells ( n = 5 mice per group ) among CD45 + cells were analyzed by fluorescence - activated cell sorting ( FACS ) and represented as box plots Data information : data are presented by Box plots Min to Max showing all points with median . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 and * * * * p < 0 . 0001 ; Mann - Whitney test .

O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

A B16F10 melanoma cells with either mTRF2 overexpression or knockdown were inoculated into the backs of immunocompetent mice. The percentages of intratumoral CD11bHi GR1Hi MDSCs (n = 10 mice), CD25+ Foxp3+ regulatory T cells (Tregs), or CD25+ Foxp3− activated T cells (n = 5 mice per group) among CD45+ cells were analyzed by fluorescence-activated cell sorting (FACS) and represented as box plots Data information: data are presented by Box plots Min to Max showing all points with median. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001; Mann-Whitney test.


TRF2 - overexpressing or - knockdown BJcl2 cells were inoculated into the backs of immunodeficient mice in the presence of Matrigel . Five days later , the percentage of immune cells infiltrating the Matrigel plugs of CD11bHi GR1Hi MDSCs ( C ) were determined by FACS ( n = 5 mice per group ) .

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TRF2-overexpressing or -knockdown BJcl2 cells were inoculated into the backs of immunodeficient mice in the presence of Matrigel. Five days later, the percentage of immune cells infiltrating the Matrigel plugs of CD11bHi GR1Hi MDSCs (C) were determined by FACS (n = 5 mice per group).


TRF2 - overexpressing or - knockdown BJcl2 cells were inoculated into the backs of immunodeficient mice in the presence of Matrigel . Five days later , the intranuclear staining of pSTAT3 ( D ) were determined by FACS ( n = 5 mice per group ) .

B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

TRF2-overexpressing or -knockdown BJcl2 cells were inoculated into the backs of immunodeficient mice in the presence of Matrigel. Five days later , the intranuclear staining of pSTAT3 (D) were determined by FACS (n = 5 mice per group).


TRF2 - overexpressing or - knockdown BJcl2 cells were inoculated into the backs of immunodeficient mice in the presence of Matrigel . Five days later , CD3 − NKp46 + and IFN - γ + among CD3 − NKp46 + NK cells ( E ) were determined by FACS ( n = 5 mice per group ) .

B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O

TRF2-overexpressing or -knockdown BJcl2 cells were inoculated into the backs of immunodeficient mice in the presence of Matrigel. Five days later, CD3− NKp46+ and IFN-γ+ among CD3− NKp46+ NK cells (E) were determined by FACS (n = 5 mice per group).


Linear correlation plot of the percentage of IFN - γ + NK cells , the mean of fluorescence intensity ( MFI ) of pSTAT3 for MDSCs , and the expression of TRF2 ( r2 = 0 , 5177 ; Pearson ' s correlation , p = 0 . 003 ) ( F ) Data information : data are presented by Box plots Min to Max showing all points with median . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 and * * * * p < 0 . 0001 ; Mann - Whitney test . Pearson correlation and p values are indicated .

O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Linear correlation plot of the percentage of IFN- γ + NK cells, the mean of fluorescence intensity (MFI) of pSTAT3 for MDSCs, and the expression of TRF2 (r2= 0,5177; Pearson's correlation, p = 0.003) (F) Data information: data are presented by Box plots Min to Max showing all points with median. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001; Mann-Whitney test. Pearson correlation and p values are indicated.


the mRNA expression level in the matrigel plugs of arg1 , pSTAT3 MFI and TRF2 expression ( r2 = 0 , 7077 ; Pearson ' s correlation , p < 0 . 0001 ) ( G ) ; the mRNA expression level in the matrigel plugs of tgfβ , pSTAT3 MFI and TRF2 expression ( r2 = 0 , 3669 ; Pearson ' s correlation , p = 0 . 0046 ) ( H ) and the mRNA expression level in the matrigel plugs of il - 10 , pSTAT3 MFI and TRF2 expression ( r2 = 0 , 7418 ; Pearson ' s correlation , p < 0 . 0001 ) ( I ) . Data information : data are presented by Box plots Min to Max showing all points with median . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 and * * * * p < 0 . 0001 ; Mann - Whitney test . Pearson correlation and p values are indicated .

O O O O O O O O O B-MEASURED_VAR O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

the mRNA expression level in the matrigel plugs of arg1, pSTAT3 MFI and TRF2 expression (r2= 0,7077; Pearson's correlation, p < 0.0001) (G); the mRNA expression level in the matrigel plugs of tgfβ, pSTAT3 MFI and TRF2 expression (r2= 0,3669; Pearson's correlation, p = 0.0046) (H) and the mRNA expression level in the matrigel plugs of il-10, pSTAT3 MFI and TRF2 expression (r2= 0,7418; Pearson's correlation, p < 0.0001) (I). Data information: data are presented by Box plots Min to Max showing all points with median. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001; Mann-Whitney test. Pearson correlation and p values are indicated.


B16F10 melanoma cells with or without mTRF2 overexpression were inoculated into the backs of immunocompetent mice and treated with anti - GR1 antibody or isotypic control ( 200µg / IP ) . The tumor volume was determined every 2 days using hemi - ellipsoid formula ( B ) . Tumor volume are presented in mm3 along the time in days . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B16F10 melanoma cells with or without mTRF2 overexpression were inoculated into the backs of immunocompetent mice and treated with anti-GR1 antibody or isotypic control (200µg/IP). The tumor volume was determined every 2 days using hemi-ellipsoid formula (B). Tumor volume are presented in mm3 along the time in days. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


B16F10 melanoma cells with or without mTRF2 overexpression were inoculated into the backs of immunocompetent mice and treated with anti - GR1 antibody or isotypic control ( 200µg / IP ) . On day 19 of the tumor growth experiment , the percentages of GFP + cells infiltrating the lungs were determined by FACS and presented as the number of GFP + cells / mg lung tissue ( C ) . Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B16F10 melanoma cells with or without mTRF2 overexpression were inoculated into the backs of immunocompetent mice and treated with anti-GR1 antibody or isotypic control (200µg/IP). On day 19 of the tumor growth experiment, the percentages of GFP+ cells infiltrating the lungs were determined by FACS and presented as the number of GFP+ cells/mg lung tissue (C). Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


On day 21 , lung metastasis was analyzed by counting macroscopic metastatic nodules ( E scale bars = 500µM .

On day 21, lung metastasis was analyzed by counting macroscopic metastatic nodules (E scale bars = 500µM.


B16F10 melanoma cells with or without mTRF2 overexpression were inoculated into tail vein of immunocompetent mice and treated with anti - GR1 antibody or isotypic control ( 200µg / IP ) every 3 days . On day 21 , lung metastasis was analyzed by hematoxylin and eosin ( H & E ) staining F ) , scale bars = 500µM . Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

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B16F10 melanoma cells with or without mTRF2 overexpression were inoculated into tail vein of immunocompetent mice and treated with anti-GR1 antibody or isotypic control (200µg/IP) every 3 days. On day 21, lung metastasis was analyzed by hematoxylin and eosin (H&E) staining F), scale bars = 500µM. Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


Tumor - infiltrating immune cells from tumors were analyzed by FACS : MDSCs and CD25 + Foxp3 + Tregs ( G ) Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

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Tumor-infiltrating immune cells from tumors were analyzed by FACS: MDSCs and CD25+ Foxp3+ Tregs (G) Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


Tumor - infiltrating immune cells from tumors were analyzed by FACS NK cells ( H ) Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

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Tumor-infiltrating immune cells from tumors were analyzed by FACS NK cells (H) Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


Tumor - infiltrating immune cells from tumors were analyzed by FACS : CD8 + T cells ( I ) . Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

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Tumor-infiltrating immune cells from tumors were analyzed by FACS: CD8+ T cells (I). Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


BJcl2 cells with or without hTRF2 overexpression were inoculated into the backs of immunodeficient Nude mice and treated with anti - GR1 antibody or isotypic control ( 200µg / IP ) and NK cells recruitment , degranulation and MDSC infiltration were monitored by FACS Density plot of CD11bHi GR1Hi MDSC staining in control or anti - GR1 - treated Matrigel plugs ( K ) .

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BJcl2 cells with or without hTRF2 overexpression were inoculated into the backs of immunodeficient Nude mice and treated with anti-GR1 antibody or isotypic control (200µg/IP) and NK cells recruitment, degranulation and MDSC infiltration were monitored by FACS Density plot of CD11bHi GR1Hi MDSC staining in control or anti-GR1-treated Matrigel plugs (K).


BJcl2 cells with or without hTRF2 overexpression were inoculated into the backs of immunodeficient Nude mice and treated with anti - GR1 antibody or isotypic control ( 200µg / IP ) and NK cells recruitment , degranulation and MDSC infiltration were monitored by FACS Box plots of the percentages of infiltrating or activated NK cells ( CD107a or CD69 ) ( L ) Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

BJcl2 cells with or without hTRF2 overexpression were inoculated into the backs of immunodeficient Nude mice and treated with anti-GR1 antibody or isotypic control (200µg/IP) and NK cells recruitment, degranulation and MDSC infiltration were monitored by FACS Box plots of the percentages of infiltrating or activated NK cells (CD107a or CD69) (L) Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


BJcl2 cells with or without hTRF2 overexpression were inoculated into the backs of immunodeficient Nude mice and treated with anti - GR1 antibody or isotypic control ( 200µg / IP ) and NK cells recruitment , degranulation and MDSC infiltration were monitored by FACS Box plots of the percentages of CD11bHi GR1Hi MDSC or NKp46 + NK cells ( M ) infiltrating the Matrigel plugs assay among the CD45 + cells . Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + cells or the indicated parent population or the number of lung nodules . All experiments are performed with n = 8 mice per group ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Mann - Whitney test .

O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O B-MEASURED_VAR O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

BJcl2 cells with or without hTRF2 overexpression were inoculated into the backs of immunodeficient Nude mice and treated with anti-GR1 antibody or isotypic control (200µg/IP) and NK cells recruitment, degranulation and MDSC infiltration were monitored by FACS Box plots of the percentages of CD11bHi GR1Hi MDSC or NKp46+ NK cells (M) infiltrating the Matrigel plugs assay among the CD45+ cells. Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ cells or the indicated parent population or the number of lung nodules. All experiments are performed with n = 8 mice per group; *p < 0.05, **p < 0.01, and ***p < 0.001; Mann-Whitney test.


Analysis of STAT3 phosphorylation by MSC2 cells after 18 hours ' co - culture with BJcl2 overexpressing or compromised for TRF2 Histograms of the pSTAT3 mean fluorescence intensity ( MFI ) ( B ) .

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Analysis of STAT3 phosphorylation by MSC2 cells after 18 hours' co-culture with BJcl2 overexpressing or compromised for TRF2 Histograms of the pSTAT3 mean fluorescence intensity (MFI) (B).


+ / - inhibitors : anti - MyD88 peptide inhibitor , anti - mouse TLR2 antibody , or lipopolysaccharide ( LPS ) - RS , an antagonist of TLR4 in ( D )

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+/- inhibitors: anti-MyD88 peptide inhibitor, anti-mouse TLR2 antibody, or lipopolysaccharide (LPS)-RS, an antagonist of TLR4 in (D)


+ / - inhibitors : anti - mouse IL - 6 or JAK1 / 2 inhibitor ( Ruxolitinib ) in ( E ) .

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+/- inhibitors: anti-mouse IL-6 or JAK1/2 inhibitor (Ruxolitinib) in (E).


TRF2 - overexpressing or - knockdown BJcl2 cells were co - cultured with MSC2 cells for 18h + / - mouse IL6 blocking antibody or Ruxolitinib . After FACS sorting , MSC2 cells were co - cultured for 18h with primed and purified NK cells before a 4h challenge with YAK - 1 . NK cell degranulation and IFN - γ production were analyzed by FACS ( G ) . Data information : Panels show all points with mean ± SEM corresponding to the percentages of each cell population among NK cells ( G ) ( n = 3 independent experiments ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Student ' s t - test ) .

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TRF2-overexpressing or -knockdown BJcl2 cells were co-cultured with MSC2 cells for 18h +/- mouse IL6 blocking antibody or Ruxolitinib. After FACS sorting, MSC2 cells were co-cultured for 18h with primed and purified NK cells before a 4h challenge with YAK-1. NK cell degranulation and IFN-γ production were analyzed by FACS (G). Data information: Panels show all points with mean ± SEM corresponding to the percentages of each cell population among NK cells (G) (n = 3 independent experiments; *p < 0.05, **p < 0.01, and ***p < 0.001; Student's t-test).


TRF2 - overexpressing or - knockdown BJcl2 cells were co - cultured with MSC2 cells for 18h + / - mouse IL6 blocking antibody or Ruxolitinib . Alternatively , a 4h challenge with NIH3T3 cells was performed and the ability of NK cells to kill NIH3T3 cells were determined by colorimetric analysis ( AlamarBlue ) ( H ) . Data information : Panels show all points with mean ± SEM corresponding to the percentage of specific target lysis ( H ) ( n = 3 independent experiments ; * p < 0 . 05 , * * p < 0 . 01 , and * * * p < 0 . 001 ; Student ' s t - test ) .

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TRF2-overexpressing or -knockdown BJcl2 cells were co-cultured with MSC2 cells for 18h +/- mouse IL6 blocking antibody or Ruxolitinib. Alternatively, a 4h challenge with NIH3T3 cells was performed and the ability of NK cells to kill NIH3T3 cells were determined by colorimetric analysis (AlamarBlue) (H). Data information: Panels show all points with mean ± SEM corresponding to the percentage of specific target lysis (H) (n = 3 independent experiments; *p < 0.05, **p < 0.01, and ***p < 0.001; Student's t-test).


Gene expression analysis by quantitative polymerase chain reaction ( qPCR ) of TRF2 target genes following TRF2 overexpression or knockdown . Data information : data represent n = 3 independent experiments with mean ±

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Gene expression analysis by quantitative polymerase chain reaction (qPCR) of TRF2 target genes following TRF2 overexpression or knockdown. Data information: data represent n = 3 independent experiments with mean ± SEM


B , C Heatmap of H3K27ac ( B ) or H3K23me3 ( C ) peaks on the 4 kilobase ( Kb ) region around genomic ITSs ( ENCODE data for the K562 cell line ) . The mean intensity of the region is depicted in the histograms . ITSs are clustered depending of the peak mean intensity obtained from H3K27ac or H3K27me3 chromatin immunoprecipitation sequencing ( ChIP - seq ) of the 4 Kb region . ITSs detected by ChIP - seq using an anti - TRF2 antibody are shown on the left .

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B, C Heatmap of H3K27ac (B) or H3K23me3 (C) peaks on the 4 kilobase (Kb) region around genomic ITSs (ENCODE data for the K562 cell line). The mean intensity of the region is depicted in the histograms. ITSs are clustered depending of the peak mean intensity obtained from H3K27ac or H3K27me3 chromatin immunoprecipitation sequencing (ChIP-seq) of the 4 Kb region. ITSs detected by ChIP-seq using an anti-TRF2 antibody are shown on the left.


D Specific qPCR for HS3SH4 , GPC6 , and VCAN ITSs after ChIP using an anti - H3K27ac antibody in TRF2 - overexpressing or - knockdown BJcl2 cells . Data represent fold enrichment of the H3K27ac level compared with the total H3 level . Data information : data represent n = 3 independent experiments with mean ±

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D Specific qPCR for HS3SH4, GPC6, and VCAN ITSs after ChIP using an anti-H3K27ac antibody in TRF2-overexpressing or -knockdown BJcl2 cells. Data represent fold enrichment of the H3K27ac level compared with the total H3 level. Data information: data represent n = 3 independent experiments with mean ± SEM


B - E Percentage of CD11bHi GR1Hi MDSCs ( B ) , CD3 - NKp46 + NK cells ( C ) , NKp46 + CD107a + NK cells ( D ) or CD69 + NK cells ( E ) Data information : data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45 + or NK cells ( n = 8 mice per group ; * p < 0 . 05 and * * * p < 0 . 001 ; Mann - Whitney test ) .

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B-E Percentage of CD11bHi GR1Hi MDSCs (B), CD3- NKp46+ NK cells (C), NKp46+ CD107a+ NK cells (D) or CD69+ NK cells (E) Data information: data are presented by Box plots Min to Max showing all points with median corresponding to the percentages of each cell population among CD45+ or NK cells (n = 8 mice per group; *p < 0.05 and ***p < 0.001; Mann-Whitney test).


Histograms of the pSTAT3 mean fluorescence intensity ( MFI ) of MSC2 cells when TRF2 was overexpressed and the target gene knocked down ( G ) Data information MFI of n = 3 independent experiment is normalized to controls and represented as fold change of MFI ( n = 3 independent experiments ; * p < 0 . 05 and * * p < 0 . 01 ; Student ' s t - test ) .

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Histograms of the pSTAT3 mean fluorescence intensity (MFI) of MSC2 cells when TRF2 was overexpressed and the target gene knocked down (G) Data information MFI of n=3 independent experiment is normalized to controls and represented as fold change of MFI (n = 3 independent experiments; *p < 0.05 and **p < 0.01; Student's t-test).


Histograms of the pSTAT3 mean fluorescence intensity ( MFI ) of MSC2 cells when both TRF2 and its target gene were knocked down ( H ) . Data information : MFI of n = 3 independent experiment is normalized to controls and represented as fold change of MFI ( n = 3 independent experiments ; * p < 0 . 05 and * * p < 0 . 01 ; Student ' s t - test ) .

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Histograms of the pSTAT3 mean fluorescence intensity (MFI) of MSC2 cells when both TRF2 and its target gene were knocked down (H). Data information: MFI of n=3 independent experiment is normalized to controls and represented as fold change of MFI (n = 3 independent experiments; *p < 0.05 and **p < 0.01; Student's t-test).


I Histograms of tumor weights on day 19 after implantation of subcutaneous xenografts containing BJcl2 cells overexpressing or knocked down for HS3ST4 or GPC6 Data information : data are represented as mean + / - SEM ( n = 8 mice per group ; * p < 0 . 05 , and * * p < 0 . 01 ; Mann - Whitney test ) .

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I Histograms of tumor weights on day 19 after implantation of subcutaneous xenografts containing BJcl2 cells overexpressing or knocked down for HS3ST4 or GPC6 Data information: data are represented as mean +/- SEM (n = 8 mice per group; *p < 0.05, and **p < 0.01; Mann-Whitney test).


A Analysis of Heparan Sulfate ( HS ) expression by FACS using the 10E4 antibody . Data information histograms representative ( left panels ) and MFIs from three independent experiments ( * p < 0 . 05 and * * p < 0 . 01 ; Student ' s t - test ) are shown + / - SEM .

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A Analysis of Heparan Sulfate (HS) expression by FACS using the 10E4 antibody. Data information histograms representative (left panels) and MFIs from three independent experiments (*p < 0.05 and **p < 0.01; Student's t-test) are shown +/- SEM.


B Atomic Force Microscopic ( AFM ) analysis of glycocalyx and cell stiffness and glycocalyx length . Data information : In ( B ) , Box plots Min to Max showing all points with median represent four independent experiments , each performed using at least 20 cells ( * p < 0 . 05 ; Mann - Whitney test ) .

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B Atomic Force Microscopic (AFM) analysis of glycocalyx and cell stiffness and glycocalyx length. Data information: In (B), Box plots Min to Max showing all points with median represent four independent experiments, each performed using at least 20 cells (*p< 0.05; Mann-Whitney test).


C AFM images showing the Modulus heatmap ( stiffness ) in GFP - expressing or TRF2 - overexpressing cancer cells . At each of the 128x128 force curve points , corresponding to an area of 10 μm2 , the AFM software automatically displays the Sneddon modulus analysis while scanning the sample . The color bar on the left of the images shows a qualitative representation of the elastic map .

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C AFM images showing the Modulus heatmap (stiffness) in GFP-expressing or TRF2-overexpressing cancer cells. At each of the 128x128 force curve points, corresponding to an area of 10 μm2, the AFM software automatically displays the Sneddon modulus analysis while scanning the sample. The color bar on the left of the images shows a qualitative representation of the elastic map.


D Analysis of pSTAT3 levels in MDSCs after co - culture with GFP - expressing or TRF2 - overexpressing cancer cells treated with or without sodium chlorate ( NaClO3 ) . Data information : In ( D ) , data represents mean of n = 3 independent experiments + / - SEM ( * * * p < 0 . 001 ; Student ' s t - test ) .

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D Analysis of pSTAT3 levels in MDSCs after co-culture with GFP-expressing or TRF2-overexpressing cancer cells treated with or without sodium chlorate (NaClO3). Data information: In (D), data represents mean of n = 3 independent experiments +/- SEM (***p < 0.001; Student's t-test).


A Kaplan - Meier curves for overall survival of chemotherapy - treated patients according to the TRF2 expression level Data information : the optimal cut - off is determined on KMplot . The p value ( log Rank test ) , the Hazard Ratio and number of patients are indicated .

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A Kaplan-Meier curves for overall survival of chemotherapy-treated patients according to the TRF2 expression level Data information: the optimal cut-off is determined on KMplot. The p value (log Rank test), the Hazard Ratio and number of patients are indicated.


Immunocompetent mice were subcutaneously engrafted with B16F10 cells with or without TRF2 overexpression and treated with three intraperitoneal injections of 50 mg / kg 5 - FU . The tumor volume was determined every 2 days using a caliper , and growth curves over time ( C ) Data information : , data represent mean values + / - SEM ( n = 8 mice per group ; * p < 0 . 05 and * * p < 0 . 01 ; Mann - Whitney test ) .

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Immunocompetent mice were subcutaneously engrafted with B16F10 cells with or without TRF2 overexpression and treated with three intraperitoneal injections of 50 mg/kg 5-FU. The tumor volume was determined every 2 days using a caliper, and growth curves over time (C) Data information: , data represent mean values +/- SEM (n = 8 mice per group; *p < 0.05 and **p < 0.01; Mann-Whitney test).


Immunocompetent mice were subcutaneously engrafted with B16F10 cells with or without TRF2 overexpression and treated with three intraperitoneal injections of 50 mg / kg 5 - FU . tumor volume were determined on day 19 and represented using Box plots Min to Max showing all points with median ( D ) . Data information : ( D ) , data are represented by Box plots Min to Max showing all points with median ( n = 8 mice per group ; * p < 0 . 05 and * * p < 0 . 01 ; Mann - Whitney test )

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Immunocompetent mice were subcutaneously engrafted with B16F10 cells with or without TRF2 overexpression and treated with three intraperitoneal injections of 50 mg/kg 5-FU. tumor volume were determined on day 19 and represented using Box plots Min to Max showing all points with median (D). Data information: (D), data are represented by Box plots Min to Max showing all points with median (n = 8 mice per group; *p < 0.05 and **p < 0.01; Mann-Whitney test)


Immunocompetent mice were subcutaneously engrafted with B16F10 cells with or without TRF2 overexpression and treated with three intraperitoneal injections of 50 mg / kg 5 - FU . Overall survival was also determined ( E ) .

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Immunocompetent mice were subcutaneously engrafted with B16F10 cells with or without TRF2 overexpression and treated with three intraperitoneal injections of 50 mg/kg 5-FU. Overall survival was also determined (E).


A Kaplan - Meier curves for overall survival of patients at all stages of cancers ( breast , gastric , ovarian , and lung cancer ) depending on TRF2 expression level . Data information : For all panels , the optimal cut - off is determined on KMplot . The p value ( log Rank test ) , the Hazard Ratio and number of patients are indicated .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

A Kaplan-Meier curves for overall survival of patients at all stages of cancers (breast, gastric, ovarian, and lung cancer) depending on TRF2 expression level. Data information: For all panels, the optimal cut-off is determined on KMplot. The p value (log Rank test), the Hazard Ratio and number of patients are indicated.


The overall survival of gastric cancer patients ( all stages ) was analyzed depending on TRF2 and its targets genes expression level ( HS3ST4 , GPC6 and VCAN collectively named the HSPG genes ) Data information : For all panels , the optimal cut - off is determined on KMplot . The p value ( log Rank test ) , the Hazard Ratio and number of patients are indicated .

O O O O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

The overall survival of gastric cancer patients (all stages) was analyzed depending on TRF2 and its targets genes expression level (HS3ST4, GPC6 and VCAN collectively named the HSPG genes) Data information: For all panels, the optimal cut-off is determined on KMplot. The p value (log Rank test), the Hazard Ratio and number of patients are indicated.


The overall survival of gastric cancer patients ( all stages ) was analyzed depending on TRF2 and with MDSC level ( Lo or Hi ) , determine as the mean expression level of CD33 and C5AR , using KMplot Data information : For all panels , the optimal cut - off is determined on KMplot . The p value ( log Rank test ) , the Hazard Ratio and number of patients are indicated .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

The overall survival of gastric cancer patients (all stages) was analyzed depending on TRF2 and with MDSC level (Lo or Hi), determine as the mean expression level of CD33 and C5AR, using KMplot Data information: For all panels, the optimal cut-off is determined on KMplot. The p value (log Rank test), the Hazard Ratio and number of patients are indicated.


E Repartition of number of gastric cancer patients depending on TRF2 and MDSC classification . Data information : For all panels , the optimal cut - off is determined on KMplot . The p value ( log Rank test ) , the Hazard Ratio and number of patients are indicated .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

E Repartition of number of gastric cancer patients depending on TRF2 and MDSC classification. Data information: For all panels, the optimal cut-off is determined on KMplot. The p value (log Rank test), the Hazard Ratio and number of patients are indicated.


F Overall survival of gastric cancer patients ( all stages ) analyzed depending on TRF2 , HSPG genes ( HS3ST4 , GPC6 and VCAN ) and MDSC level ( Lo or Hi ) . Data information : For all panels , the optimal cut - off is determined on KMplot . The p value ( log Rank test ) , the Hazard Ratio and number of patients are indicated .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

F Overall survival of gastric cancer patients (all stages) analyzed depending on TRF2, HSPG genes (HS3ST4, GPC6 and VCAN) and MDSC level (Lo or Hi). Data information: For all panels, the optimal cut-off is determined on KMplot. The p value (log Rank test), the Hazard Ratio and number of patients are indicated.


Testing an assay for SARS - CoV - 2 using synthetic RNA targets . We show data from both fluorescent ( a ) and lateral flow detection ( b ) . Target concentrations ( in cp / μl ) are indicated . RPA NTC : water input into RPA ; Det NTC : water input into Cas13 detection reaction . In ( a ) , error bars indicate one standard deviation based on n = 3 technical replicates .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Testing an assay for SARS-CoV-2 using synthetic RNA targets. We show data from both fluorescent (a) and lateral flow detection (b). Target concentrations (in cp/μl) are indicated. RPA NTC: water input into RPA; Det NTC: water input into Cas13 detection reaction. In (a), error bars indicate one standard deviation based on n=3 technical replicates.


mRNA expression of ANG in colonic tissues from normal controls ( n = 25 ) and UC ( n = 27 ) or CD ( n = 46 ) patients .

O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O

mRNA expression of ANG in colonic tissues from normal controls (n = 25) and UC (n = 27) or CD (n = 46) patients.


Representative western blotting of ANG in colonic tissues from normal controls and UC or CD patients .

O O O O B-MEASURED_VAR O O O O O O O O O O O O

Representative western blotting of ANG in colonic tissues from normal controls and UC or CD patients.


Representative images showing ANG immunohistochemical ( IHC ) staining in normal control and UC or CD sample .

O O O B-MEASURED_VAR O O O O O O O O O O O O O O

Representative images showing ANG immunohistochemical (IHC) staining in normal control and UC or CD sample.


Corresponding statistical analysis of ANG expression score in normal controls ( n = 10 ) and UC ( n = 10 ) or CD ( n = 15 ) patients .

O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O

Corresponding statistical analysis of ANG expression score in normal controls (n = 10) and UC (n = 10) or CD (n = 15) patients.


Representative images showing hematoxylin - eosin ( HE ) staining and ANG IHC staining in mild and severe IBD samples .

O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O

Representative images showing hematoxylin-eosin (HE) staining and ANG IHC staining in mild and severe IBD samples.


Statistical analyses of ANG expression scores in mild ( n = 18 ) and severe ( n = 19 ) UC ( F ) or mild ( n = 20 ) and severe ( n = 20 ) CD ( G ) patients .

O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Statistical analyses of ANG expression scores in mild (n = 18) and severe (n = 19) UC (F) or mild (n = 20) and severe (n = 20) CD (G) patients.


Ang mRNA expression in colonic tissue from 2 . 5 % DSS - treated WT mice at indicated time point ( n = 4 mice / time point ) . DSS : dextran sulfate

B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Ang mRNA expression in colonic tissue from 2.5% DSS-treated WT mice at indicated time point (n = 4 mice/time point). DSS: dextran sulfate sodium


Kaplan - Meier curve of 3 . 5 % DSS - treated Ang - deficient mice ( Ang - / - , n = 14 ) or littermate controls ( WT , n = 13 ) .

O O O O O O O O O O O O B-CONTROLLED_VAR O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O

Kaplan-Meier curve of 3.5% DSS-treated Ang-deficient mice (Ang-/-, n = 14) or littermate controls (WT, n = 13).


Body weight loss of WT and Ang - / - mice with ( n = 9 ) or without ( n = 6 ) 2 . 5 % DSS treatment .

O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O

Body weight loss of WT and Ang-/- mice with (n = 9) or without (n = 6) 2.5% DSS treatment.


disease activity index ( DAI ) of WT and Ang - / - mice with ( n = 9 ) or without ( n = 6 ) 2 . 5 % DSS treatment .

O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O

disease activity index (DAI) of WT and Ang-/- mice with (n = 9) or without (n = 6) 2.5% DSS treatment.


Colon length of colonic section from the WT and Ang - / - mice on day 9 with ( n = 9 ) or without ( n = 6 ) 2 . 5 % DSS treatment .

O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O

Colon length of colonic section from the WT and Ang-/- mice on day 9 with (n = 9) or without (n = 6) 2.5% DSS treatment.


serum FITC - Dextran level of colonic section from the WT and Ang - / - mice on day 9 with ( n = 9 ) or without ( n = 6 ) 2 . 5 % DSS treatment . FITC - Dextran : fluorescein isothiocyanate - labeled dextran .

O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

serum FITC-Dextran level of colonic section from the WT and Ang-/- mice on day 9 with (n = 9) or without (n = 6) 2.5% DSS treatment. FITC-Dextran: fluorescein isothiocyanate-labeled dextran.


representative HE staining image , and ( H ) histopathological score of colonic section from the WT and Ang - / - mice on day 9 with ( n = 9 ) or without ( n = 6 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O

representative HE staining image, and (H) histopathological score of colonic section from the WT and Ang-/- mice on day 9 with (n = 9) or without (n = 6) 2.5% DSS treatment.


Quantitative mRNA expression of cytokine genes in colonic tissue from the WT and Ang - / - mice on day 0 ( n = 3 ) or day 9 ( n = 9 ) during 2 . 5 % DSS treatment .

O O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Quantitative mRNA expression of cytokine genes in colonic tissue from the WT and Ang-/- mice on day 0 (n = 3) or day 9 (n = 9) during 2.5% DSS treatment.


Soluble cytokine level in supernatant of cultured colonic tissue isolated from WT and Ang - / - mice on day 0 ( n = 3 ) or day 9 ( n = 7 ) during 2 . 5 % DSS treatment .

Soluble cytokine level in supernatant of cultured colonic tissue isolated from WT and Ang-/- mice on day 0 (n = 3) or day 9 (n = 7) during 2.5% DSS treatment.


Body weight loss of WT → WT , Ang - / - → WT and WT → Ang - / - chimera mice with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O B-CONTROLLED_VAR O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O

Body weight loss of WT→WT, Ang-/-→WT and WT→Ang-/- chimera mice with (n = 7) or without (n = 5) 2.5% DSS treatment.


DAI of WT → WT , Ang - / - → WT and WT → Ang - / - chimera mice with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O B-CONTROLLED_VAR O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O

DAI of WT→WT, Ang-/-→WT and WT→Ang-/- chimera mice with (n = 7) or without (n = 5) 2.5% DSS treatment.


Colon length of colonic section from these chimera colitis mice on day 8 with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Colon length of colonic section from these chimera colitis mice on day 8 with (n = 7) or without (n = 5) 2.5% DSS treatment.


serum FITC - Dextran level , of colonic section from these chimera colitis mice on day 8 with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

serum FITC-Dextran level, of colonic section from these chimera colitis mice on day 8 with (n = 7) or without (n = 5) 2.5% DSS treatment.


representative HE staining image , and ( F ) histopathological score of colonic section from these chimera colitis mice on day 8 with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

representative HE staining image, and (F) histopathological score of colonic section from these chimera colitis mice on day 8 with (n = 7) or without (n = 5) 2.5% DSS treatment.


Ang mRNA expression in mouse lymphoid and myeloid cells ; data represent two independent experiments ( n = 2 ) .

B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O

Ang mRNA expression in mouse lymphoid and myeloid cells; data represent two independent experiments (n = 2).


Body weight loss of Angfl / fl and LyzMcre ; Angfl / fl mice with ( n = 7 ) or without ( n = 3 ) 2 . 5 % DSS treatment .

O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O

Body weight loss of Angfl/fl and LyzMcre;Angfl/fl mice with (n = 7) or without (n = 3) 2.5% DSS treatment.


DAI of Angfl / fl and LyzMcre ; Angfl / fl mice with ( n = 7 ) or without ( n = 3 ) 2 . 5 % DSS treatment .

O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O

DAI of Angfl/fl and LyzMcre;Angfl/fl mice with (n = 7) or without (n = 3) 2.5% DSS treatment.


Colon length of colonic section from the Angfl / fl and LyzMcre ; Angfl / fl mice on day 8 with ( n = 7 ) or without ( n = 3 ) 2 . 5 % DSS treatment .

O O O O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O

Colon length of colonic section from the Angfl/fl and LyzMcre;Angfl/fl mice on day 8 with (n = 7) or without (n = 3) 2.5% DSS treatment.


serum FITC - Dextran level , of colonic section from the Angfl / fl and LyzMcre ; Angfl / fl mice on day 8 with ( n = 7 ) or without ( n = 3 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O

serum FITC-Dextran level, of colonic section from the Angfl/fl and LyzMcre;Angfl/fl mice on day 8 with (n = 7) or without (n = 3) 2.5% DSS treatment.


representative HE staining image , and ( M ) histopathological score of colonic section from the Angfl / fl and LyzMcre ; Angfl / fl mice on day 8 with ( n = 7 ) or without ( n = 3 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O

representative HE staining image, and (M) histopathological score of colonic section from the Angfl/fl and LyzMcre;Angfl/fl mice on day 8 with (n = 7) or without (n = 3) 2.5% DSS treatment.


Quantitative mRNA expression of ANG and PLXNB2 in isolated IECs or colonic laminar propria ( cLP ) cells from WT mice ( n = 9 ) .

O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O

Quantitative mRNA expression of ANG and PLXNB2 in isolated IECs or colonic laminar propria (cLP) cells from WT mice (n = 9).


western blotting of ANG and PLXNB2 in isolated IECs or colonic laminar propria ( cLP ) cells from WT mice ( n = 9 ) .

O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O

western blotting of ANG and PLXNB2 in isolated IECs or colonic laminar propria (cLP) cells from WT mice (n = 9).


Representative images showing immunofluorescence staining of ANG , PLXNB2 and macrophage surface marker F4 / 80 in frozen colonic section from WT mice .

O O O O O O B-MEASURED_VAR O B-MEASURED_VAR O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O

Representative images showing immunofluorescence staining of ANG, PLXNB2 and macrophage surface marker F4/80 in frozen colonic section from WT mice.


Representative images showing immunofluorescence staining of GFP and PLXNB2 in frozen colonic section from WT mice infected with adeno - associated virus ( AAV9 ) .

O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O

Representative images showing immunofluorescence staining of GFP and PLXNB2 in frozen colonic section from WT mice infected with adeno-associated virus (AAV9).


PLXNB2 expression in IECs from the AAV9 - infected WT mice ( n = 3 ) .

B-MEASURED_VAR O O O O O O O O O O O O O O O O

PLXNB2 expression in IECs from the AAV9-infected WT mice (n = 3).


Body weight loss of the AA9 - infected WT mice with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Body weight loss of the AA9-infected WT mice with (n = 7) or without (n = 5) 2.5% DSS treatment.


DAI of the AA9 - infected WT mice with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O

DAI of the AA9-infected WT mice with (n = 7) or without (n = 5) 2.5% DSS treatment.


Colon length of colonic section from the AAV9 - infected WT mice on day 8 with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Colon length of colonic section from the AAV9-infected WT mice on day 8 with (n = 7) or without (n = 5) 2.5% DSS treatment.


serum FITC - Dextran level of colonic section from the AAV9 - infected WT mice on day 8 with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

serum FITC-Dextran level of colonic section from the AAV9-infected WT mice on day 8 with (n = 7) or without (n = 5) 2.5% DSS treatment.


representative HE staining image , and ( K ) histopathological score of colonic section from the AAV9 - infected WT mice on day 8 with ( n = 7 ) or without ( n = 5 ) 2 . 5 % DSS treatment .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

representative HE staining image, and (K) histopathological score of colonic section from the AAV9-infected WT mice on day 8 with (n = 7) or without (n = 5) 2.5% DSS treatment.


Representative images showing cleaved - caspase 3 staining in colonic section from WT or Ang - / - colitis mice ( n = 3 ) at indicated time point ; arrows indicate cleaved - caspase 3 positive cells ; two fields per colon section are quantified .

O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O

Representative images showing cleaved-caspase 3 staining in colonic section from WT or Ang-/- colitis mice (n = 3) at indicated time point; arrows indicate cleaved-caspase 3 positive cells; two fields per colon section are quantified.


the number of apoptotic cells in colonic section from WT or Ang - / - colitis mice ( n = 3 ) at indicated time

O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O

the number of apoptotic cells in colonic section from WT or Ang-/- colitis mice (n = 3) at indicated time point


Cell apoptotic status of IECs from WT or Ang - / - colitis mice ( n = 5 ) on day 4 .

O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O

Cell apoptotic status of IECs from WT or Ang-/- colitis mice (n = 5) on day 4.


Representative images showing Ki67 staining in colonic section from WT or Ang - / - colitis mice ( n = 3 ) at indicated time point ; five representative crypts per colon section are quantified .

O O O B-MEASURED_VAR O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O

Representative images showing Ki67 staining in colonic section from WT or Ang-/- colitis mice (n = 3) at indicated time point; five representative crypts per colon section are quantified.


the number of proliferating cells per crypt in colonic section from WT or Ang - / - colitis mice ( n = 3 ) at indicated time point ; five representative crypts per colon section are quantified .

O O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O

the number of proliferating cells per crypt in colonic section from WT or Ang-/- colitis mice (n = 3) at indicated time point; five representative crypts per colon section are quantified.


Cell - cycle status of IECs from WT or Ang - / - colitis mice ( n = 5 ) on day 4 .

O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O

Cell-cycle status of IECs from WT or Ang-/- colitis mice (n = 5) on day 4.


Representative images showing cleaved - caspase 3 staining in colonic section from Angfl / fl and LyzMCre ; Angfl / fl , or AAV - shControl - and AAV - shPlxnb2 - treated colitis mice on day 4 ( n = 3 ) ; arrows indicate cleaved - caspase 3 positive cells ; two fields per colon section are quantified .

O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O

Representative images showing cleaved-caspase 3 staining in colonic section from Angfl/fl and LyzMCre;Angfl/fl, or AAV-shControl- and AAV-shPlxnb2- treated colitis mice on day 4 (n = 3); arrows indicate cleaved-caspase 3 positive cells; two fields per colon section are quantified.


the number of apoptotic cells in colonic section from Angfl / fl and LyzMCre ; Angfl / fl , or AAV - shControl - and AAV - shPlxnb2 - treated colitis mice on day 4 ( n = 3 )

O O O O O O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O

the number of apoptotic cells in colonic section from Angfl/fl and LyzMCre;Angfl/fl, or AAV-shControl- and AAV-shPlxnb2- treated colitis mice on day 4 (n = 3)


Ki67 staining and ( L ) the number of proliferating cells per crypt in representative colonic section from Angfl / fl and LyzMCre ; Angfl / fl , or AAV - shControl - and AAV - shPlxnb2 - treated colitis mice on day 4 ( n = 3 ) ; five representative crypts per colon section are quantified .

B-MEASURED_VAR O O O O O O O O O O O O O O O O O B-CONTROLLED_VAR O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

Ki67 staining and (L) the number of proliferating cells per crypt in representative colonic section from Angfl/fl and LyzMCre;Angfl/fl, or AAV-shControl- and AAV-shPlxnb2- treated colitis mice on day 4 (n = 3); five representative crypts per colon section are quantified.


PLXNB2 expressions in HCT116 ( including wild type HCT116 ( HCT116WT ) and ANG knockout HCT116 ( HCT116KO ) ) and THP - 1 as detected by flow cytometry analysis . The upper three dash lines represent the intensity of unstained negative control of these cells , while the solid lines represent the intensity of PLXNB2 antibody staining .

B-MEASURED_VAR O O O O O O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O

PLXNB2 expressions in HCT116 (including wild type HCT116 (HCT116WT) and ANG knockout HCT116 (HCT116KO)) and THP-1 as detected by flow cytometry analysis. The upper three dash lines represent the intensity of unstained negative control of these cells, while the solid lines represent the intensity of PLXNB2 antibody staining.


Secreted ANG level in conditioned medium from THP1 , HCT116WT or HCT116KO ( n = 3 ) .

O B-MEASURED_VAR O O O O O O O O O O O O O O O O

Secreted ANG level in conditioned medium from THP1, HCT116WT or HCT116KO (n = 3).


The interaction between ANGWT , ANGR66A or ANGK40I with PLXNB2 in HCT116KO detected by co - immunoprecipitation assay . ANGR66A , ANG receptor binding site variant ; ANGK40I , ANG enzymatic activity null

O O O B-MEASURED_VAR O B-MEASURED_VAR O B-MEASURED_VAR O B-MEASURED_VAR O O O O O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O

The interaction between ANGWT, ANGR66A or ANGK40I with PLXNB2 in HCT116KO detected by co-immunoprecipitation assay. ANGR66A, ANG receptor binding site variant; ANGK40I, ANG enzymatic activity null variant


THP - 1 - derived ANG translocating into HCT116KO through PLXNB2 in co - cultured system as showed by western blotting . Neamine , a blocker of ANG and PLXNB2 interaction ; siCtrl : control siRNA ; siPLXNB2 - 1 and siPLXNB2 -

O O O O O B-MEASURED_VAR O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O O O O O

THP-1-derived ANG translocating into HCT116KO through PLXNB2 in co-cultured system as showed by western blotting. Neamine, a blocker of ANG and PLXNB2 interaction; siCtrl: control siRNA; siPLXNB2-1 and siPLXNB2-2


Flow cytometry analysis of HCT116KO stained with Annexin V and PI following different treatments as indicated . ANGR66A , ANG receptor binding site variant ; ANGK40I , ANG enzymatic activity null variant ; BAY , inhibitor of TNF - α - induced κB kinase ( IKK )

O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O B-CONTROLLED_VAR I-CONTROLLED_VAR I-CONTROLLED_VAR O O B-CONTROLLED_VAR I-CONTROLLED_VAR O B-CONTROLLED_VAR O

Flow cytometry analysis of HCT116KO stained with Annexin V and PI following different treatments as indicated. ANGR66A, ANG receptor binding site variant; ANGK40I, ANG enzymatic activity null variant; BAY, inhibitor of TNF-α-induced κB kinase (IKK) phosphorylation


tiRNA and 5 ' - tiRNAAla production in HCT116KO following different treatments as indicated . ANGR66A , ANG receptor binding site variant ; ANGK40I , ANG enzymatic activity null variant ; BAY , inhibitor of TNF - α - induced κB kinase ( IKK )

B-MEASURED_VAR O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O B-CONTROLLED_VAR I-CONTROLLED_VAR I-CONTROLLED_VAR O O B-CONTROLLED_VAR I-CONTROLLED_VAR O B-CONTROLLED_VAR O

tiRNA and 5'-tiRNAAla production in HCT116KO following different treatments as indicated. ANGR66A, ANG receptor binding site variant; ANGK40I, ANG enzymatic activity null variant; BAY, inhibitor of TNF-α-induced κB kinase (IKK) phosphorylation


The effect of 5 ' - tiRNA knockdown or overexpression on TNF - α - induced apoptosis in ANGWT - or ANGK40I - treated HCT116KO . si5 ' - tiRNA : tiRNA - targeting

O O O B-CONTROLLED_VAR I-CONTROLLED_VAR I-CONTROLLED_VAR I-CONTROLLED_VAR O O O O O O O O O O O B-CONTROLLED_VAR O O B-CONTROLLED_VAR O O O O O B-CONTROLLED_VAR I-CONTROLLED_VAR I-CONTROLLED_VAR O B-CONTROLLED_VAR O O

The effect of 5'-tiRNA knockdown or overexpression on TNF-α-induced apoptosis in ANGWT- or ANGK40I-treated HCT116KO. si5'-tiRNA: tiRNA-targeting siRNA


The interaction between 5 ' - tiRNA and cytochrome c ( Cyt c ) in HCT116 cells as detected by RIP .

O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O

The interaction between 5'-tiRNA and cytochrome c (Cyt c) in HCT116 cells as detected by RIP.


Caspase activity of HCT116KO following different treatments as indicated . ANGR66A , ANG receptor binding site variant ; ANGK40I , ANG enzymatic activity null variant ; BAY , inhibitor of TNF - α - induced κB kinase ( IKK )

O O O O O O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O B-CONTROLLED_VAR O B-CONTROLLED_VAR O O O O O O O O O B-CONTROLLED_VAR I-CONTROLLED_VAR I-CONTROLLED_VAR O O B-CONTROLLED_VAR I-CONTROLLED_VAR O B-CONTROLLED_VAR O

Caspase activity of HCT116KO following different treatments as indicated. ANGR66A, ANG receptor binding site variant; ANGK40I, ANG enzymatic activity null variant; BAY, inhibitor of TNF-α-induced κB kinase (IKK) phosphorylation


Cell viability of HCT116KO following different treatments as indicated . siCtrl : control siRNA ; siPLXNB2 - 1 and siPLXNB2 - 2 : PLXNB2

O O O O O O O O O O O O O O O O O O O O O O O B-CONTROLLED_VAR

Cell viability of HCT116KO following different treatments as indicated. siCtrl: control siRNA; siPLXNB2-1 and siPLXNB2-2: PLXNB2 siRNAs


Detection of 47S pre - rRNA transcription level with northern blotting in HCT116KO following different treatments as indicated . siPLXNB2 - 1 and siPLXNB2 - 2 : PLXNB2

O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O B-CONTROLLED_VAR

Detection of 47S pre-rRNA transcription level with northern blotting in HCT116KO following different treatments as indicated. siPLXNB2-1 and siPLXNB2-2: PLXNB2 siRNAs


tiRNA and 5 ' - tiRNAAla production in IECs isolated from WT and Ang - / - colitis mice at indicated time point .

B-MEASURED_VAR O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O

tiRNA and 5'-tiRNAAla production in IECs isolated from WT and Ang-/- colitis mice at indicated time point.


47S pre - rRNA transcription level in IECs from WT and Ang - / - colitis mice at indicated time point .

B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O B-CONTROLLED_VAR O O O O O O O O O O

47S pre-rRNA transcription level in IECs from WT and Ang-/- colitis mice at indicated time point.


Western blotting of ANG in colonic tissue from WT mice ( n = 2 ) at indicated time point in the ANG prevention model . The grayscale of ANG band in WT mice with no recombinant ANG and DSS treatment ( untreated ) is arbitrarily set as 1 .

O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O B-MEASURED_VAR O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O

Western blotting of ANG in colonic tissue from WT mice (n = 2) at indicated time point in the ANG prevention model. The grayscale of ANG band in WT mice with no recombinant ANG and DSS treatment (untreated) is arbitrarily set as 1.


Body weight loss of the pre - treated WT colitis mice ( n = 8 ) .

Body weight loss of the pre-treated WT colitis mice (n = 8).


DAI of the pre - treated WT colitis mice ( n = 8 ) .

DAI of the pre-treated WT colitis mice (n = 8).


Colon length of colon section from the pre - treated WT colitis mice on day 8 ( n = 8 ) .

O O O O O O O O O O O O O O O O O O O O O O

Colon length of colon section from the pre-treated WT colitis mice on day 8 (n = 8).


serum FITC - dextran level of colon section from the pre - treated WT colitis mice on day 8 ( n = 8 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O

serum FITC-dextran level of colon section from the pre-treated WT colitis mice on day 8 (n = 8).


representative HE staining image , and ( H ) histopathological score of colon section from the pre - treated WT colitis mice on day 8 ( n = 8 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

representative HE staining image, and (H) histopathological score of colon section from the pre-treated WT colitis mice on day 8 (n = 8).


Western blotting of ANG in colonic tissue from WT ( n = 2 ) mice at indicated time point in the ANG treatment model . The grayscale of ANG band in WT mice with no recombinant ANG and DSS treatment ( untreated ) is arbitrarily set as 1 .

Western blotting of ANG in colonic tissue from WT (n = 2) mice at indicated time point in the ANG treatment model. The grayscale of ANG band in WT mice with no recombinant ANG and DSS treatment (untreated) is arbitrarily set as 1.


Body weight loss of the treated WT colitis mice ( n = 8 ) .

Body weight loss of the treated WT colitis mice (n = 8).


DAI of the treated WT colitis mice ( n = 8 ) .

DAI of the treated WT colitis mice (n = 8).


Colon length of colon sections from the treated WT colitis mice on day 8 ( n = 8 ) .

Colon length of colon sections from the treated WT colitis mice on day 8 (n = 8).


serum FITC - dextran level of colon sections from the treated WT colitis mice on day 8 ( n = 8 ) .

O O O O O O O O O O O O O O O O O O O O O O O

serum FITC-dextran level of colon sections from the treated WT colitis mice on day 8 (n = 8).


representative HE staining image , and ( P ) histopathological score of colon sections from the treated WT colitis mice on day 8 ( n = 8 ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

representative HE staining image, and (P) histopathological score of colon sections from the treated WT colitis mice on day 8 (n = 8).


B . Top : representative Western blots assessing mTOR phosphorylation at S2448 and p70S6K phosphorylation at T389 as indexes of mTOR activity in PFC of adult WT mice are illustrated . Bottom : data represent the ratios of immunoreactive signals of the anti phospho - mTOR ( S2448 ) or anti phospho - p70S6K ( T389 ) antibodies to the immunoreactive signal of the anti - β - actin antibody and are expressed in % of values in vehicle - injected WT mice . They are the means ± S . E . M . of results obtained in five mice per group . * p < 0 . 05 ; * * p < 0 . 01 , one - way ANOVA followed by Newman - Keuls test .

O O O O O O O O B-MEASURED_VAR O O O O B-MEASURED_VAR O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B. Top: representative Western blots assessing mTOR phosphorylation at S2448 and p70S6K phosphorylation at T389 as indexes of mTOR activity in PFC of adult WT mice are illustrated. Bottom: data represent the ratios of immunoreactive signals of the anti phospho-mTOR (S2448) or anti phospho-p70S6K (T389) antibodies to the immunoreactive signal of the anti-β-actin antibody and are expressed in % of values in vehicle-injected WT mice. They are the means ± S.E.M. of results obtained in five mice per group. * p < 0.05; ** p < 0.01, one-way ANOVA followed by Newman-Keuls test.


C . Top : schemas illustrating the behavioral tasks examined in WT mice . Bottom : the plots represent the discrimination index for the novel object recognition task ( Vehicle N = 11 , THC : N = 11 , THC + SB : N = 11 , THC + Rapa : N = 12 ) , the sociability index ( Vehicle : N = 11 , THC : N = 11 , THC + SB : N = 11 , THC + Rapa : N = 12 ) and the discrimination index for the social discrimination task ( N = 8 for each group ) , measured in each condition . * p < 0 . 05 , * * p < 0 . 01 , one - way ANOVA followed by Bonferroni test ( error bars correspond to the mean ± SEM , the dotted line to a discrimination index ( exploration of novel object - exploration of familiar object / total object exploration ) equal to zero .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

C. Top: schemas illustrating the behavioral tasks examined in WT mice. Bottom: the plots represent the discrimination index for the novel object recognition task (Vehicle N=11, THC: N=11, THC+SB: N=11, THC+Rapa: N=12), the sociability index (Vehicle: N=11, THC: N=11, THC+SB: N=11, THC+Rapa: N=12) and the discrimination index for the social discrimination task (N=8 for each group), measured in each condition. * p < 0.05, ** p < 0.01, one-way ANOVA followed by Bonferroni test (error bars correspond to the mean ± SEM, the dotted line to a discrimination index (exploration of novel object - exploration of familiar object/total object exploration) equal to zero.


D . Top : representative Western blots assessing mTOR phosphorylation at S2448 and p70S6K phosphorylation at T389 in PFC of adult 5 - HT6 - / - mice are illustrated . Bottom : data represent the ratios of immunoreactive signals of the anti phospho - mTOR ( S2448 ) or anti phospho - p70S6K ( T389 ) antibodies to the immunoreactive signal of the anti - β - actin antibody and are expressed in % of values in vehicle - injected 5 - HT6 - / - mice . They are the means ± S . E . M . of results obtained in four mice per group . n . s . p > 0 . 05 , one - way ANOVA followed by Newman - Keuls test .

O O O O O O O O B-MEASURED_VAR O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

D. Top: representative Western blots assessing mTOR phosphorylation at S2448 and p70S6K phosphorylation at T389 in PFC of adult 5-HT6-/- mice are illustrated. Bottom: data represent the ratios of immunoreactive signals of the anti phospho-mTOR (S2448) or anti phospho-p70S6K (T389) antibodies to the immunoreactive signal of the anti-β-actin antibody and are expressed in % of values in vehicle-injected 5-HT6-/- mice. They are the means ± S.E.M. of results obtained in four mice per group. n.s. p > 0.05, one-way ANOVA followed by Newman-Keuls test.


E . Top : schemas illustrating the behavioral tasks in 5 - HT6 - / - mice . Bottom : the plots represent the discrimination index for the novel object recognition test ( discrimination index : 0 . 30 ± 0 . 05 , N = 10 and 0 . 31 ± 0 . 03 , N = 11 , for THC + Vehicle and Vehicle + Vehicle conditions , respectively , p > 0 . 05 ) , the 3 - chamber social preference test ( sociability index : 0 . 49 ± 0 . 04 , N = 10 and 0 . 48 ± 0 . 07 , N = 11 for THC + Vehicle and Vehicle + Vehicle conditions , respectively , p > 0 . 05 ) and the social discrimination test ( discrimination index : 0 . 15 ± 0 . 06 , N = 8 and 0 . 21 ± 0 . 06 , N = 8 for THC + Vehicle and Vehicle + Vehicle conditions , respectively , p > 0 . 05 ) , measured in each condition . One - way ANOVA followed by Bonferroni test ( error bars correspond to the mean ± SEM , the dotted line corresponds to a discrimination index equal to zero ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

E. Top: schemas illustrating the behavioral tasks in 5-HT6-/- mice. Bottom: the plots represent the discrimination index for the novel object recognition test (discrimination index: 0.30 ± 0.05, N=10 and 0.31 ± 0.03, N=11, for THC+Vehicle and Vehicle+Vehicle conditions, respectively, p > 0.05), the 3-chamber social preference test (sociability index: 0.49 ± 0.04, N=10 and 0.48 ± 0.07, N=11 for THC+Vehicle and Vehicle+Vehicle conditions, respectively, p > 0.05) and the social discrimination test (discrimination index: 0.15 ± 0.06, N=8 and 0.21 ± 0.06, N=8 for THC+Vehicle and Vehicle+Vehicle conditions, respectively, p > 0.05), measured in each condition. One-way ANOVA followed by Bonferroni test (error bars correspond to the mean ± SEM, the dotted line corresponds to a discrimination index equal to zero).


A - E . The PFC was stained with CB1 receptor ( A1 - D1 , green ) or 5HT6 receptor ( E1 - H1 , green ) , Bassoon ( A2 and E2 , red ) , GAD65 ( B2 and F2 , red ) , PSD - 95 ( C2 and G2 , red ) , or SERT ( D2 and H2 , red ) antibodies . Merge images are also depicted ( A3 - H3 ) . CB1 receptors were mainly co - localized with the presynaptic marker Bassoon ( white arrows , A3 ) , mostly within GABAergic boutons ( white arrows , B3 ) and to a lesser extend with PSD - 95 ( white arrows , C3 ) and SERT ( D3 ) , while 5 - HT6 receptors were mainly co - localized with PSD - 95 ( white arrows , G3 ) . No co - localization of CB1 and 5 - HT6 receptors was detected ( white arrows indicate the 5 - HT6 staining , H3 ) . Scale bar : 10 μm . The scatter plot represents the co - localization analysis for CB1 and 5 - HT6 receptor immunostainings . Graph on the top , Mander ' s split coefficient was used to identify the fraction of CB1 receptors that co - localizes with Bassoon ( Mander ' s coefficient : 0 . 66 ± 0 . 02 , n = 4 ) , GAD65 ( Mander ' s coefficient : 0 . 56 ± 0 . 01 , n = 4 ) , PSD95 ( Mander ' s coefficient : 0 . 15 ± 0 . 01 , n = 4 ) or SERT ( Mander ' s coefficient : 0 . 082 ± 0 . 004 , n = 4 ) . Graph on the bottom . Mander ' s split coefficient was used to identify the fraction of 5 - HT6 receptors that co - localizes with Bassoon ( Mander ' s coefficient : 0 . 09 ± 0 . 01 , n = 4 ) , GAD65 ( Mander ' s coefficient : 0 . 059 ± 0 . 004 , n = 4 ) , PSD95 ( Mander ' s coefficient : 0 . 58 ± 0 . 01 , n = 4 ) or CB1 receptors ( Mander ' s coefficient : 0 . 0020 ± 0 . 0004 , n = 4 ) . Error bars correspond to the mean ± SEM .

O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O B-MEASURED_VAR O O O O O O O O O B-MEASURED_VAR O O O O O O O O B-MEASURED_VAR O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O B-MEASURED_VAR O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR I-MEASURED_VAR I-MEASURED_VAR O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

A-E. The PFC was stained with CB1 receptor (A1-D1, green) or 5HT6 receptor (E1-H1, green), Bassoon (A2 and E2, red), GAD65 (B2 and F2, red), PSD-95 (C2 and G2, red), or SERT (D2 and H2, red) antibodies. Merge images are also depicted (A3-H3). CB1 receptors were mainly co-localized with the presynaptic marker Bassoon (white arrows, A3), mostly within GABAergic boutons (white arrows, B3) and to a lesser extend with PSD-95 (white arrows, C3) and SERT (D3), while 5-HT6 receptors were mainly co-localized with PSD-95 (white arrows, G3). No co-localization of CB1 and 5-HT6 receptors was detected (white arrows indicate the 5-HT6 staining, H3). Scale bar: 10 μm. The scatter plot represents the co-localization analysis for CB1 and 5-HT6 receptor immunostainings. Graph on the top, Mander's split coefficient was used to identify the fraction of CB1 receptors that co-localizes with Bassoon (Mander's coefficient: 0.66 ± 0.02, n=4), GAD65 (Mander's coefficient: 0.56 ± 0.01, n=4), PSD95 (Mander's coefficient: 0.15 ± 0.01, n=4) or SERT (Mander's coefficient: 0.082 ± 0.004, n=4). Graph on the bottom. Mander's split coefficient was used to identify the fraction of 5-HT6 receptors that co-localizes with Bassoon (Mander's coefficient: 0.09 ± 0.01, n=4), GAD65 (Mander's coefficient: 0.059 ± 0.004, n=4), PSD95 (Mander's coefficient: 0.58 ± 0.01, n=4) or CB1 receptors (Mander's coefficient: 0.0020 ± 0.0004, n=4). Error bars correspond to the mean ± SEM.


B . Top : representative Western blots assessing mTOR activity in PFC are illustrated . Bottom : data represent the ratios of immunoreactive signals of the anti phospho - mTOR ( S2448 ) or anti phospho - p70S6K ( T389 ) antibodies to the immunoreactive signal of the anti - β - actin antibody and are expressed in % of values in vehicle - injected mice . They are the means ± S . E . M . of results obtained in six mice per group . * p < 0 . 05 ; * * p < 0 . 01 , one - way ANOVA followed by Newman - Keuls test .

O O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O B-MEASURED_VAR O O O O O O O B-MEASURED_VAR O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

B. Top: representative Western blots assessing mTOR activity in PFC are illustrated. Bottom: data represent the ratios of immunoreactive signals of the anti phospho-mTOR (S2448) or anti phospho-p70S6K (T389) antibodies to the immunoreactive signal of the anti-β-actin antibody and are expressed in % of values in vehicle-injected mice. They are the means ± S.E.M. of results obtained in six mice per group. * p < 0.05; ** p < 0.01, one-way ANOVA followed by Newman-Keuls test.


C . The plots represent the discrimination index measured in each condition . * * * p < 0 . 001 , one - way ANOVA followed by Bonferroni test . The discrimination index for the novel object recognition task is 0 . 34 ± 0 . 06 ( N = 14 ) and - 0 . 08 ± 0 . 07 ( N = 20 ) , for mice injected with Vehicle and THC , respectively , p < 0 . 001 and - 0 . 09 ± 0 . 08 ( N = 21 ) and 0 . 00 ± 0 . 06 ( N = 18 ) , for mice treated with THC + SB258285 and THC + Rapa , respectively , p > 0 . 05 vs . THC - injected mice ( error bars correspond to the mean ± SEM , the dotted line to a discrimination index equal to zero ) .

O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

C. The plots represent the discrimination index measured in each condition. *** p < 0.001, one-way ANOVA followed by Bonferroni test. The discrimination index for the novel object recognition task is 0.34 ± 0.06 (N=14) and -0.08 ± 0.07 (N= 20), for mice injected with Vehicle and THC, respectively, p < 0.001 and -0.09 ± 0.08 (N=21) and 0.00 ± 0.06 (N=18), for mice treated with THC+SB258285 and THC+Rapa, respectively, p > 0.05 vs. THC-injected mice (error bars correspond to the mean ± SEM, the dotted line to a discrimination index equal to zero).