( H ) . Parasitemia of cultures of iRBCs challenged with neutrophils in presence or absence of catalase .

O O O O B-EXP_ASSAY O O O B-CELL_TYPE O O B-CELL_TYPE O O O O O B-SMALL_MOLECULE O

(H). Parasitemia of cultures of iRBCs challenged with neutrophils in the presence or absence of catalase.


( I ) . Short - term luciferase - killing using neutrophils from three different donors in presence or absence of 5 uM cytochalasin D .

O O O O O O O B-GENEPROD O B-EXP_ASSAY O B-CELL_TYPE O O O O O O O O O O O B-SMALL_MOLECULE I-SMALL_MOLECULE O

(I). Short-term luciferase-based killing assay using neutrophils from three different donors in the presence or absence of 5 uM cytochalasin D.


( A ) . Representative flow cytometric of human neutrophils cultured alone ( Control ) , incubated with late - stage NF54 ( wild type ) or PfEMP1 Knock out ( PfEMP1 KO ) GFP + iRBC .

O O O O O B-EXP_ASSAY I-EXP_ASSAY O B-ORGANISM B-CELL_TYPE O O O O O O O O O O O B-ORGANISM O O O O O B-ORGANISM O O O B-ORGANISM O O B-GENEPROD O B-CELL_TYPE O

(A). Representative flow cytometric analysis of human neutrophils cultured alone (Control), incubated with late-stage NF54 parasites (wild type) or PfEMP1 Knock out (PfEMP1 KO) GFP+ iRBC.


( B ) . Flow cytometric of effect of trypsin treatment on neutrophil interaction with wild type PfEMP1 KO GFP + iRBC .

O O O O B-EXP_ASSAY I-EXP_ASSAY O O O B-SMALL_MOLECULE O O B-CELL_TYPE O O O O B-ORGANISM O B-GENEPROD O B-CELL_TYPE O

(B). Flow cytometric quantification of the effect of trypsin treatment on neutrophil interaction with wild type and PfEMP1 KO GFP+ iRBC.


( C ) . Luciferase - killing neutrophil elimination of late - stage wild type ( wt ) PfEMP1ko iRBC .

O O O O B-GENEPROD O B-EXP_ASSAY B-CELL_TYPE O O O O O O O O O O B-ORGANISM B-CELL_TYPE O

(C). Luciferase-based killing assay and neutrophil elimination of late-stage wild type (wt) and PfEMP1ko iRBC.


( B ) . Short - term ( 6h ) neutrophil killing of unselected NF54 antibody - selected PFD1235w expressing lines .

O O O O O O O O O O B-CELL_TYPE O O O B-ORGANISM O O O O O O O

(B). Short-term (6h) neutrophil killing of unselected NF54 and antibody-selected PFD1235w expressing parasite lines.


( C ) . Flow cytometric of neutrophil interaction with MitoTracker ( APC + ) stained NF54 iRBCs antibody - selected PFD1235w expressing iRBC .

(C). Flow cytometric quantification of neutrophil interaction with MitoTracker (APC+) stained NF54 iRBCs and antibody-selected PFD1235w expressing iRBC.


( D ) . reduction in parasitemia of NF54 antibody - selected PFD1235w expressing after 5 days of co - culture with neutrophils , compared to unchallenged

O O O O O O B-EXP_ASSAY O B-ORGANISM O O O O O O O O O O O O O B-CELL_TYPE O O O O

(D). Percent reduction in parasitemia of NF54 and antibody-selected PFD1235w expressing parasites after 5 days of co-culture with neutrophils, compared to unchallenged parasites


( E ) . var gene transcription profiles measured by qRT - PCR of antibody - selected PFD1235w expressing cultured in absence ( upper panel ) or presence ( lower panel ) of neutrophils . Steady state mRNA of each var gene was calculated as in A . I .

O O O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O O O O O O O O O O O O O O O O O O O O B-CELL_TYPE O O O O O O O O O O O O O O O O

(E). var gene transcription profiles measured by qRT-PCR of antibody-selected PFD1235w expressing parasite line cultured in the absence (upper panel) or presence (lower panel) of neutrophils. Steady state mRNA levels of each individual var gene was calculated as in A. I.


( A ) QRT - PCR of PAX6 , SOX1 , NKX2 - 1 , NKX2 - 2 , TBR1 TBR2 during dorsal ( up ) ventral ( down ) neural differentiation . ( B ) QRT - PCR of SOX1 - OT V1 during dorsal ( up ) ventral ( down ) neural differentiation Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 ( unpaired two - Student ' s t - ) .

O O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O B-GENEPROD O B-GENEPROD O B-GENEPROD I-GENEPROD I-GENEPROD O B-GENEPROD I-GENEPROD I-GENEPROD O B-GENEPROD I-GENEPROD O B-TISSUE O O O B-TISSUE O O O B-CELL_TYPE O O O O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O B-GENEPROD O O O O B-TISSUE O O O B-TISSUE O O O B-CELL_TYPE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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) QRT-PCR assay of PAX6, SOX1, NKX2-1, NKX2-2, TBR1 and TBR2 during dorsal (up) and ventral (down) neural differentiation. (B) QRT-PCR assay of SOX1-OT V1 during dorsal (up) and ventral (down) neural differentiation Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 (unpaired two-tailed Student's t-test).


( F G ) Immunostaining of PAX6 ( green ) SOX1 ( red ) in control V1 - PAKI on day 16 of dorsal neural differentiation ( F ) , quantiﬁcation of SOX1 + ( G ) , p = 0 . 0001 . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O B-GENEPROD O O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and G) Immunostaining assay of PAX6 (green) and SOX1 (red) in control and V1-PAKI cells on day 16 of dorsal neural differentiation (F), and quantiﬁcation of SOX1+ cells (G), p=0.0001. Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 (unpaired two-tailed Student's t-test).


( H I ) Immunostaining of MAP2 ( green ) in control V1 - PAKI on day 35 of dorsal neural differentiation ( H ) , quantiﬁcation of MAP2 + ( I ) , p = 0 . 0001 . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and I) Immunostaining assay of MAP2 (green) in control and V1-PAKI cells on day 35 of dorsal neural differentiation (H), and quantiﬁcation of MAP2+ cells (I), p=0.0001. Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 (unpaired two-tailed Student's t-test).


( J K ) Immunostaining of PAX6 ( green ) SOX1 ( red ) in control V1 - PAKI on day 16 of ventral neural differentiation ( J ) , quantiﬁcation of SOX1 + ( K ) , p = 0 . 0003 . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 ( unpaired two - Student ' s t - ) .

(J and K) Immunostaining assay of PAX6 (green) and SOX1 (red) in control and V1-PAKI cells on day 16 of ventral neural differentiation (J), and quantiﬁcation of SOX1+ cells (K), p=0.0003. Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 (unpaired two-tailed Student's t-test).


( L M ) Immunostaining of GABA ( green ) in control V1 - PAKI on day 35 of ventral neural differentiation ( L ) , quantiﬁcation of GABA + ( M ) , p = 0 . 0001 . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O B-SMALL_MOLECULE O O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

(L and M) Immunostaining assay of GABA (green) in control and V1-PAKI cells on day 35 of ventral neural differentiation (L), and quantiﬁcation of GABA+ cells (M), p=0.0001. Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 (unpaired two-tailed Student's t-test).


( C D ) Immunostaining of MAP2 ( green ) in control SOX1KO on day 35 of dorsal neural differentiation ( C ) , quantiﬁcation of MAP2 + ( D ) , p = 0 . 0001 . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * * * p < 0 . 001 versus Ctrl group ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O O B-GENEPROD O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and D) Immunostaining assay of MAP2 (green) in control and SOX1KO cells on day 35 of dorsal neural differentiation (C), and quantiﬁcation of MAP2+ cells (D), p=0.0001. Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). ***p<0.001 versus Ctrl group (unpaired two-tailed Student's t-test).


( F G ) Immunostaining of GABA ( green ) in control SOX1KO on day 35 of ventral neural differentiation ( F ) , quantiﬁcation of GABA + ( G ) , p = 0 . 0001 . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * * * p < 0 . 001 versus Ctrl group ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O B-SMALL_MOLECULE O O O O O B-GENEPROD O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and G) Immunostaining assay of GABA (green) in control and SOX1KO cells on day 35 of ventral neural differentiation (F), and quantiﬁcation of GABA+ cells (G), p=0.0001. Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). ***p<0.001 versus Ctrl group (unpaired two-tailed Student's t-test).


( H I ) Immunostaining of MAP2 ( green ) after SOX1 overexpression in V1 - PAKI on day 35 of dorsal neural differentiation ( H ) , quantiﬁcation of MAP2 + ( I ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( V1 - PAKI + ptSOX1 + dox ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * * * p < 0 . 001 versus Ctrl group ; # # # p < 0 . 001 versus - dox group ( one - way ANOVA ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O 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-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O 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-SMALL_MOLECULE O O O O O O O O

(H and I) Immunostaining assay of MAP2 (green) after SOX1 overexpression in V1-PAKI cells on day 35 of dorsal neural differentiation (H), and quantiﬁcation of MAP2+ cells (I), p=0.0001 (V1-PAKI), p=0.0001 (V1-PAKI+ptSOX1+dox). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). ***p<0.001 versus Ctrl group ; ###p<0.001 versus -dox group (one-way ANOVA).


( J K ) Immunostaining of GABA ( green ) after SOX1 overexpression in V1 - PAKI on day 35 of ventral neural differentiation ( J ) , quantiﬁcation of GABA + ( K ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0002 ( V1 - PAKI + ptSOX1 + dox ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * * * p < 0 . 001 versus Ctrl group ; # # # p < 0 . 001 versus - dox group ( one - way ANOVA ) .

O O O O B-EXP_ASSAY O B-SMALL_MOLECULE O O O O B-GENEPROD O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-SMALL_MOLECULE O O O O 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-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O 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-SMALL_MOLECULE O O O O O O O O

(J and K) Immunostaining assay of GABA (green) after SOX1 overexpression in V1-PAKI cells on day 35 of ventral neural differentiation (J), and quantiﬁcation of GABA+ cells (K), p=0.0001 (V1-PAKI), p=0.0002 (V1-PAKI+ptSOX1+dox). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). ***p<0.001 versus Ctrl group ; ###p<0.001 versus -dox group (one-way ANOVA).


( A B ) Enrichment of H3K14Ac ( left ) H3K27Ac ( right ) on SOX1 promoter region in control V1 - PAKI on day 16 of dorsal neural differentiation ( A ) , p = 0 . 0046 ( H3K14Ac ) , p = 0 . 0223 ( H3K27Ac ) ventral neural differentiation ( B ) , p = 0 . 0499 ( H3K14Ac ) , p = 0 . 0022 ( H3K27Ac ) . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 versus Ctrl group ( unpaired two - Student ' s t - ) .

O O O O O O B-GENEPROD O O O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O O O O O B-GENEPROD O O O O O O O O B-GENEPROD O B-TISSUE B-CELL_TYPE O O O O O O O O O O O B-GENEPROD O O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O O O 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 and B) Enrichment of H3K14Ac (left) and H3K27Ac (right) on SOX1 promoter region in control and V1-PAKI cells on day 16 of dorsal neural differentiation (A), p=0.0046 (H3K14Ac), p=0.0223 (H3K27Ac) and ventral neural differentiation (B), p=0.0499 (H3K14Ac), p=0.0022 (H3K27Ac). Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01 versus Ctrl group (unpaired two-tailed Student's t-test).


( C Immunostaining of SOX1 ( red ) after 4 HDAC inhibitors ( 4i ) , including RG2833 ( 2833 , 2μM ) , CAY10683 ( CAY , 2μM ) , PCI - 34051 ( 34051 , 5μM ) Bufexamac ( Buf , 12 . 5μM ) , treatment or remove one inhibitor from these 4 HDAC inhibitors ( 4i ) in V1 - PAKI on day 16 of dorsal neural differentiation Scale bar , 100 μm .

O O B-EXP_ASSAY O B-GENEPROD O O O O O B-GENEPROD O O O O O O B-SMALL_MOLECULE O B-SMALL_MOLECULE O O O O B-SMALL_MOLECULE O B-SMALL_MOLECULE O O O O B-SMALL_MOLECULE I-SMALL_MOLECULE I-SMALL_MOLECULE O B-SMALL_MOLECULE O O O B-SMALL_MOLECULE O B-SMALL_MOLECULE O O O O O O O O O O O O O O B-GENEPROD O O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O

(C Immunostaining assay of SOX1 (red) after 4 HDAC inhibitors (4i), including RG2833 (2833, 2μM), CAY10683 (CAY, 2μM), PCI-34051 (34051, 5μM) and Bufexamac (Buf, 12.5μM), treatment or remove one inhibitor from these 4 HDAC inhibitors (4i) in V1-PAKI cells on day 16 of dorsal neural differentiation Scale bar, 100 μm.


( G H ) Immunostaining of SOX1 ( red ) after Bufexamac treatment in V1 - PAKI on day 16 of dorsal neural differentiation ( G ) , quantiﬁcation of SOX1 + ( H ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( + Buf ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 versus Ctrl group ; # p < 0 . 05 , # # p < 0 . 01 , # # # p < 0 . 001 versus V1 - PAKI group ; ( one - way ANOVA ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-SMALL_MOLECULE O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O B-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

(G and H) Immunostaining assay of SOX1 (red) after Bufexamac treatment in V1-PAKI cells on day 16 of dorsal neural differentiation (G), and quantiﬁcation of SOX1+ cells (H), p=0.0001 (V1-PAKI), p=0.0001 (+Buf). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 versus Ctrl group ; #p<0.05, ##p<0.01, ###p<0.001 versus V1-PAKI group ; (one-way ANOVA).


( J K ) Immunostaining of MAP2 ( green ) after Bufexamac treatment in V1 - PAKI on day 35 of dorsal neural differentiation ( J ) , quantiﬁcation of MAP2 + ( K ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( + Buf ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 versus Ctrl group ; # p < 0 . 05 , # # p < 0 . 01 , # # # p < 0 . 001 versus V1 - PAKI group ( one - way ANOVA ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-SMALL_MOLECULE O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O B-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

(J and K) Immunostaining assay of MAP2 (green) after Bufexamac treatment in V1-PAKI cells on day 35 of dorsal neural differentiation (J), and quantiﬁcation of MAP2+ cells (K), p=0.0001 (V1-PAKI), p=0.0001 (+Buf). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 versus Ctrl group ; #p<0.05, ##p<0.01, ###p<0.001 versus V1-PAKI group (one-way ANOVA).


( L ) Enrichment of polymerase Ⅱ on SOX1 promoter region after Bufexamac treatment in V1 - PAKI on day 16 of dorsal ( left , p = 0 . 0007 ( V1 - PAKI ) , p = 0 . 0002 ( + Buf ) ) ventral ( right , p = 0 . 0083 ( V1 - PAKI ) , p = 0 . 0276 ( + Buf ) ) neural differentiation . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 versus Ctrl group ; # p < 0 . 05 , # # p < 0 . 01 , # # # p < 0 . 001 versus V1 - PAKI group ; ( one - way ANOVA ) .

O O O O O B-GENEPROD I-GENEPROD O B-GENEPROD O O O B-SMALL_MOLECULE O O O O O O O O O B-TISSUE O O O O O O O O O O O O O O O O O O O O O B-SMALL_MOLECULE O O B-TISSUE O O O O O O O O O O O O O O O O O O O O O B-SMALL_MOLECULE O O B-CELL_TYPE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

(L) Enrichment of RNA polymerase Ⅱ on SOX1 promoter region after Bufexamac treatment in V1-PAKI cells on day 16 of dorsal (left, p=0.0007 (V1-PAKI), p=0.0002 (+Buf)) and ventral (right, p=0.0083 (V1-PAKI), p=0.0276 (+Buf)) neural differentiation. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 versus Ctrl group ; #p<0.05, ##p<0.01, ###p<0.001 versus V1-PAKI group ; (one-way ANOVA).


( B C ) RIP for endogenous interaction between HDAC10 SOX1 - OT V1 on day 16 of dorsal neural differentiation ( B ) , p = 0 . 0085 ventral neural differentiation ( C ) , p = 0 . 0152 . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 versus IgG group ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O O B-EXP_ASSAY O B-GENEPROD I-GENEPROD O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and C) RIP analysis for endogenous interaction between HDAC10 and SOX1-OT V1 on day 16 of dorsal neural differentiation (B), p=0.0085 and ventral neural differentiation (C), p=0.0152. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 versus IgG group (unpaired two-tailed Student's t-test).


( D E ) Enrichment of HDAC10 on SOX1 promoter region in control V1 - PAKI on day 16 of dorsal neural differentiation ( D ) , p = 0 . 0052 ventral neural differentiation ( E ) , p = 0 . 0001 . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * p < 0 . 01 , * * * p < 0 . 001 versus Ctrl group ( unpaired two - Student ' s t - ) .

O O O O O O B-GENEPROD O B-GENEPROD O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and E) Enrichment of HDAC10 on SOX1 promoter region in control and V1-PAKI cells on day 16 of dorsal neural differentiation (D), p=0.0052 and ventral neural differentiation (E), p=0.0001. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, **p<0.01, ***p<0.001 versus Ctrl group (unpaired two-tailed Student's t-test).


( G H ) Immunostaining of SOX1 ( red ) after SOX1 - OT V1 , SOX1 - OT V1 - 5P or SOX1 - OT V1 - 3P overexpression in V1 - PAKI on day 16 of dorsal neural differentiation ( G ) , quantiﬁcation of SOX1 + ( H ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( ptV1 + dox ) , p = 0 . 0001 ( pt5P + dox ) . Scale bar , 100 μm . ( I J ) Immunostaining of SOX1 ( red ) after SOX1 - OT V1 , SOX1 - OT V1 - 5P or SOX1 - OT V1 - 3P overexpression in V1 - PAKI on day 16 of ventral neural differentiation ( I ) , quantiﬁcation of SOX1 + ( J ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( ptV1 + dox ) , p = 0 . 0001 ( pt5P + dox ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * p < 0 . 05 , * * * p < 0 . 001 versus Ctrl group ; # p < 0 . 05 , # # p < 0 . 01 , # # # p < 0 . 001 versus - dox group ( one - way ANOVA ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O B-GENEPROD O O O O O O B-GENEPROD O O O O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD 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-SMALL_MOLECULE O O O O O O O O O O B-SMALL_MOLECULE O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O B-GENEPROD O O O O O O B-GENEPROD O O O O O O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD 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-SMALL_MOLECULE O O O O O O O O O O B-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O

(G and H) Immunostaining assay of SOX1 (red) after SOX1-OT V1, SOX1-OT V1-5P or SOX1-OT V1-3P overexpression in V1-PAKI cells on day 16 of dorsal neural differentiation (G), and quantiﬁcation of SOX1+ cells (H), p=0.0001 (V1-PAKI), p=0.0001 (ptV1+dox), p=0.0001 (pt5P+dox). Scale bar, 100 μm. (I and J) Immunostaining assay of SOX1 (red) after SOX1-OT V1, SOX1-OT V1-5P or SOX1-OT V1-3P overexpression in V1-PAKI cells on day 16 of ventral neural differentiation (I), and quantiﬁcation of SOX1+ cells (J), p=0.0001 (V1-PAKI), p=0.0001 (ptV1+dox), p=0.0001 (pt5P+dox). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). *p<0.05, ***p<0.001 versus Ctrl group ; #p<0.05, ##p<0.01, ###p<0.001 versus -dox group (one-way ANOVA).


( A B ) Immunostaining of ASCL1 ( green ) in control , V1 - PAKI SOX1KO on day 25 of dorsal neural differentiation ( A ) , quantiﬁcation of ASCL1 + ( B ) , p = 0 . 0021 ( V1 - PAKI ) , p = 0 . 0024 ( SOX1KO ) . Scale bar , 100 μm . ( C D ) Immunostaining of ASCL1 ( green ) in control , V1 - PAKI SOX1KO on day 25 of ventral neural differentiation ( C ) , quantiﬁcation of ASCL1 + ( D ) , p = 0 . 0003 ( V1 - PAKI ) , p = 0 . 0004 ( SOX1KO ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * * p < 0 . 01 , * * * p < 0 . 001 versus Ctrl group ( unpaired two - Student ' s t - ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O O O O O O B-GENEPROD O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O B-GENEPROD O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O O O O O O B-GENEPROD O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O B-GENEPROD O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O 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 and B) Immunostaining assay of ASCL1 (green) in control, V1-PAKI and SOX1KO cells on day 25 of dorsal neural differentiation (A), and quantiﬁcation of ASCL1+ cells (B), p=0.0021 (V1-PAKI), p=0.0024 (SOX1KO). Scale bar, 100 μm. (C and D) Immunostaining assay of ASCL1 (green) in control, V1-PAKI and SOX1KO cells on day 25 of ventral neural differentiation (C), and quantiﬁcation of ASCL1+ cells (D), p=0.0003 (V1-PAKI), p=0.0004 (SOX1KO). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). **p<0.01, ***p<0.001 versus Ctrl group (unpaired two-tailed Student's t-test).


( E F ) Immunostaining of ASCL1 ( green ) after SOX1 overexpression in V1 - PAKI on day 25 of dorsal neural differentiation ( E ) , quantiﬁcation of ASCL1 + ( F ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( ptSOX1 + dox ) . Scale bar , 100 μm . ( G H ) Immunostaining of ASCL1 ( green ) after SOX1 overexpression in V1 - PAKI on day 25 of ventral neural differentiation ( G ) , quantiﬁcation of ASCL1 + ( H ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( ptSOX1 + dox ) . Scale bar , 100 μm . ( I J ) Immunostaining of MAP2 ( green ) after ASCL1 overexpression in V1 - PAKI on day 35 of dorsal neural differentiation ( I ) , quantiﬁcation of MAP2 + ( J ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( ptASCL1 + dox ) . Scale bar , 100 μm . ( K L ) Immunostaining of GABA ( green ) after ASCL1 overexpression in V1 - PAKI on day 35 of ventral neural differentiation ( K ) , quantiﬁcation of GABA + ( L ) , p = 0 . 0001 ( V1 - PAKI ) , p = 0 . 0001 ( ptASCL1 + dox ) . Scale bar , 100 μm . Data information : Data are presented as mean ± from three independent experiments ( n = 3 ) . * * * p < 0 . 001 versus Ctrl group ; # # # p < 0 . 001 versus - dox group ( one - way ANOVA ) .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD 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-SMALL_MOLECULE O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD 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-SMALL_MOLECULE O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-GENEPROD 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-SMALL_MOLECULE O O O O O O O O O O O O B-EXP_ASSAY O B-SMALL_MOLECULE O O O O B-GENEPROD O O O O O O O O O B-TISSUE B-CELL_TYPE O O O O O O O B-SMALL_MOLECULE 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-SMALL_MOLECULE O O O O O O O O O O O O O O O O O O O O O O O O O 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-SMALL_MOLECULE O O O O O O O O

(E and F) Immunostaining assay of ASCL1 (green) after SOX1 overexpression in V1-PAKI cells on day 25 of dorsal neural differentiation (E), and quantiﬁcation of ASCL1+ cells (F), p=0.0001 (V1-PAKI), p=0.0001 (ptSOX1+dox). Scale bar, 100 μm. (G and H) Immunostaining assay of ASCL1 (green) after SOX1 overexpression in V1-PAKI cells on day 25 of ventral neural differentiation (G), and quantiﬁcation of ASCL1+ cells (H), p=0.0001 (V1-PAKI), p=0.0001 (ptSOX1+dox). Scale bar, 100 μm. (I and J) Immunostaining assay of MAP2 (green) after ASCL1 overexpression in V1-PAKI cells on day 35 of dorsal neural differentiation (I), and quantiﬁcation of MAP2+ cells (J), p=0.0001 (V1-PAKI), p=0.0001 (ptASCL1+dox). Scale bar, 100 μm. (K and L) Immunostaining assay of GABA (green) after ASCL1 overexpression in V1-PAKI cells on day 35 of ventral neural differentiation (K), and quantiﬁcation of GABA+ cells (L), p=0.0001 (V1-PAKI), p=0.0001 (ptASCL1+dox). Scale bar, 100 μm. Data information: Data are presented as mean ±SEM from three independent experiments (n=3). ***p<0.001 versus Ctrl group ; ###p<0.001 versus -dox group (one-way ANOVA).


( A B ) Immunostaining of SOX1 ( red ) in control , V1 - PAKI SOX1KO on day 40 of dorsal ( A ) ventral ( B ) organoids differentiation . Scale bar , 100 μm . ( C ) Immunostaining of MAP2 ( green ) in control , V1 - PAKI SOX1KO on day 40 of dorsal organoids differentiation . Scale bar , 100 μm . ( D ) Immunostaining of GABA ( green ) in control , V1 - PAKI SOX1KO on day 40 of ventral organoids differentiation . Scale bar , 100 μm .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O O O O O O B-GENEPROD O O O O B-TISSUE O O O B-TISSUE O O O O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O O O O O O B-GENEPROD O O O O B-TISSUE O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O O O O O O B-GENEPROD O O O O B-TISSUE O O O O O O O O O

(A and B) Immunostaining assay of SOX1 (red) in control, V1-PAKI and SOX1KO cells on day 40 of dorsal (A) and ventral (B) organoids differentiation. Scale bar, 100 μm. (C) Immunostaining assay of MAP2 (green) in control, V1-PAKI and SOX1KO cells on day 40 of dorsal organoids differentiation. Scale bar, 100 μm. (D) Immunostaining assay of GABA (green) in control, V1-PAKI and SOX1KO cells on day 40 of ventral organoids differentiation. Scale bar, 100 μm.


( E F ) Immunostaining of SOX1 ( red ) after SOX1 - OT V1 overexpression in V1 - PAKI on day 40 of dorsal ( E ) ventral ( F ) organoids differentiation . Scale bar , 100 μm . ( G ) Immunostaining of MAP2 ( green ) after SOX1 - OT V1 overexpression in V1 - PAKI on day 40 of dorsal organoids differentiation . Scale bar , 100 μm . ( H ) Immunostaining of GABA ( green ) after SOX1 - OT V1 overexpression in V1 - PAKI on day 40 of ventral organoids differentiation . Scale bar , 100 μm .

O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O O O O B-TISSUE O O O B-TISSUE O O O O O O O O O O O O O O O B-EXP_ASSAY O B-GENEPROD O O O O B-GENEPROD O O O O O O O O O O O O B-TISSUE O O O O O O O O O O O O B-EXP_ASSAY O B-SMALL_MOLECULE O O O O B-GENEPROD O O O O O O O O O O O O B-TISSUE O O O O O O O O O

(E and F) Immunostaining assay of SOX1 (red) after SOX1-OT V1 overexpression in V1-PAKI cells on day 40 of dorsal (E) and ventral (F) organoids differentiation. Scale bar, 100 μm. (G) Immunostaining assay of MAP2 (green) after SOX1-OT V1 overexpression in V1-PAKI cells on day 40 of dorsal organoids differentiation. Scale bar, 100 μm. (H) Immunostaining assay of GABA (green) after SOX1-OT V1 overexpression in V1-PAKI cells on day 40 of ventral organoids differentiation. Scale bar, 100 μm.


B ) Total T were enriched from spleens of Ptpn6fl / fl / Ptpn11fl / flWbm , CD4cre Ptpn6fl / fl / Ptpn11fl / flWbm , CD4cre Ptpn6fl / fl mice . Expression of Ptpn6 Ptpn11 was tested by immunoblot in lysates , Gapdh used as loading control .

O O O B-CELL_TYPE O O O B-TISSUE O B-GENEPROD O O O B-GENEPROD O O O B-GENEPROD I-GENEPROD O O O B-GENEPROD O O O B-GENEPROD I-GENEPROD O O B-ORGANISM O O O B-GENEPROD I-GENEPROD O O O B-EXP_ASSAY O O O B-GENEPROD O O O O O

B) Total T cells were enriched from spleens of Ptpn6fl/fl/Ptpn11fl/flWbm, CD4cre Ptpn6fl/fl/Ptpn11fl/flWbm, CD4cre Ptpn6fl/fl mice. Expression of Ptpn6 and Ptpn11 was tested by immunoblot in cell lysates, Gapdh used as loading control.


CD4cre Ptpn6fl / fl control mice were subcutaneously injected with MC38 . Tumour growth in mice is shown for indicated genotypes treatments ; number of mice eradicating tumour is shown within graphs ( C ) .

B-GENEPROD I-GENEPROD O O O B-ORGANISM O O O O B-CELL_LINE O B-EXP_ASSAY I-EXP_ASSAY O B-ORGANISM O O O O O O O O O B-ORGANISM O O O O O O O O O O

CD4cre Ptpn6fl/fl and control mice were subcutaneously injected with MC38 cells. Tumour growth in individual mice is shown for the indicated genotypes and treatments; number of mice eradicating the tumour is shown within the graphs (C).


CD4cre Ptpn6fl / fl control mice were subcutaneously injected with MC38 . Survival curves are shown ( D ) .

B-GENEPROD I-GENEPROD O O O B-ORGANISM O O O O B-CELL_LINE O B-EXP_ASSAY I-EXP_ASSAY O O O O O O

CD4cre Ptpn6fl/fl and control mice were subcutaneously injected with MC38 cells. Survival curves are shown (D).


Tumour growth in mice challenged with MC38 subsequently treated with anti - PD - 1 antibody or isotype control is shown for each indicated genotype ; number of mice eradicating tumour is shown within graphs ( G ) .

B-EXP_ASSAY I-EXP_ASSAY O B-ORGANISM O O B-CELL_LINE O O O O O B-GENEPROD I-GENEPROD I-GENEPROD O O O O O O O O O O O O O B-ORGANISM O O O O O O O O O O

Tumour growth in individual mice challenged with MC38 and subsequently treated with anti-PD-1 antibody or isotype control is shown for each indicated genotype; number of mice eradicating the tumour is shown within the graphs (G).


mice challenged with MC38 subsequently treated with anti - PD - 1 antibody or isotype control is shown for each indicated genotype Survival curves of CD4cre Ptpn6fl / fl / Ptpn11fl / flWbm control mice is shown ( H ) .

B-ORGANISM O O B-CELL_LINE O O O O O B-GENEPROD I-GENEPROD I-GENEPROD O O O O O O O O O O B-EXP_ASSAY I-EXP_ASSAY O B-GENEPROD I-GENEPROD O O O B-GENEPROD O O O B-ORGANISM O O O O O O

individual mice challenged with MC38 and subsequently treated with anti-PD-1 antibody or isotype control is shown for each indicated genotype Survival curves of CD4cre Ptpn6fl/fl/Ptpn11fl/flWbm and control mice is shown (H).


I , J ) 10 - 12 days following MC38 tumour inoculation treatment with anti - PD - 1 antibody or isotype control , CD4cre Ptpn6 / 11wt / wt CD4cre Ptpn6fl / fl / Ptpn11fl / flWbm mice were sacrificed , TILs analysed . Graphs depict frequencies of CD8 + T ( gated as CD45 + TCRβ + CD8 + ) expressing IFN - γ TNFα upon re - stimulation ( G ) percentages of CD8 + T ( H ) .

O O O O O O O O O B-CELL_LINE O O O O O O B-GENEPROD I-GENEPROD I-GENEPROD O O O O O B-GENEPROD I-GENEPROD O O O O B-GENEPROD I-GENEPROD O O O B-GENEPROD O O B-ORGANISM O O O O O O O O O O B-CELL_TYPE I-CELL_TYPE I-CELL_TYPE O O O B-GENEPROD O B-GENEPROD O B-GENEPROD O O O B-GENEPROD I-GENEPROD I-GENEPROD I-GENEPROD O O O O O O O O O B-CELL_TYPE I-CELL_TYPE I-CELL_TYPE O O O O

I, J) 10-12 days following MC38 tumour inoculation and treatment with anti-PD-1 antibody or isotype control, CD4cre Ptpn6/11wt/wt and CD4cre Ptpn6fl/fl/Ptpn11fl/flWbm mice were sacrificed, and TILs analysed. Graphs depict frequencies of CD8+ T cells (gated as CD45+ TCRβ+ CD8+) expressing IFN-γ and TNFα upon re-stimulation (G) and percentages of CD8+ T cells (H).


B ) Ptpn6 Ptpn11 copy number , as estimated through high resolution mass spectrometry in GzmBcre Ptpn6 / 11fl / fl control CTLs .

O O B-GENEPROD I-GENEPROD B-EXP_ASSAY I-EXP_ASSAY O O O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O B-GENEPROD I-GENEPROD O O O O O B-CELL_TYPE O

B) Ptpn6 and Ptpn11 copy number, as estimated through high resolution mass spectrometry in GzmBcre Ptpn6/11fl/fl and control CTLs.


C D ) GzmBcre R26R - YFP reporter mice were injected subcutaneously with MC38 analysed after 11 days . Graphs show of YFP + among CD45 + CD8 + TILs ( C ) of PD - 1 + among CD45 + CD8 + YFP + TILs ( D ) . Results depict n = 4 mice ( C D ) .

O O O B-GENEPROD O O B-GENEPROD O B-ORGANISM O O O O B-CELL_LINE O O O O O O O O B-GENEPROD O O B-GENEPROD O B-GENEPROD O O O O O O B-GENEPROD I-GENEPROD I-GENEPROD O O B-GENEPROD O B-GENEPROD O B-GENEPROD O O O O O O O O O O O B-ORGANISM O O O O O

C and D) GzmBcre R26R-YFP reporter mice were injected subcutaneously with MC38 cells and analysed after 11 days. Graphs show the percentage of YFP+ cells among CD45+ CD8+ TILs (C) and the percentage of PD-1+ cells among CD45+ CD8+ YFP+ TILs (D). Results depict n = 4 mice (C and D).


GzmBcre Ptpn6 / 11fl / fl control Ptpn6 / 11fl / fl mice were subcutaneously injected with MC38 . Survival curves statistical comparisons between different groups are shown ( E ) .

B-GENEPROD I-GENEPROD O O O O O B-GENEPROD O O O O B-ORGANISM O O O O B-CELL_LINE O B-EXP_ASSAY I-EXP_ASSAY O O O O O O O O O O O

GzmBcre Ptpn6/11fl/fl and control Ptpn6/11fl/fl mice were subcutaneously injected with MC38 cells. Survival curves and statistical comparisons between different groups are shown (E).


tumour start depicts time ( days ) following tumour engraftment after which tumour is palpable , while tumour endpoint depicts time ( days ) from tumour start until reaching maximal allowed tumour ( F ) .

O O O O O O O O B-EXP_ASSAY I-EXP_ASSAY O O O O O O O O O O O O O O O O O O O O O B-EXP_ASSAY O O O O

The tumour start depicts the time (days) following tumour engraftment after which the tumour is palpable, while tumour endpoint depicts the time (days) from tumour start until the reaching the maximal allowed tumour size (F).


C ) Numbers of CTLs ( gated on CD8 + T ) from Ptpn6 / 11fl / fl or GzmBcre Ptpn6 / 11fl / fl mice were counted daily by flow cytometry with addition of DAPI to monitor dead .

O O O O B-CELL_TYPE O O O B-CELL_TYPE I-CELL_TYPE I-CELL_TYPE O O B-GENEPROD O O O O O B-GENEPROD I-GENEPROD O O O O B-ORGANISM O O O O B-EXP_ASSAY I-EXP_ASSAY O O O B-SMALL_MOLECULE O O O O

C) Numbers of CTLs (gated on CD8+ T cells) from Ptpn6/11fl/fl or GzmBcre Ptpn6/11fl/fl mice were counted daily by flow cytometry with the addition of DAPI to monitor dead cells.


D ) Quantitative high - resolution mass spectrometry was used to resolve proteome of Ptpn6 / 11fl / fl GzmBcre Ptpn6 / 11fl / fl CTLs ( day 7 ) . plot depicts copy number in control GzmBcre Ptpn6 / 11fl / fl . Significantly altered are indicated in red ; selected examples are annotated . Results are on n = 3 biological replicates two - , unequal - variance t - on log10 transformed copy number per values was used to compare differences between experimental groups .

O O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O O O O O O B-GENEPROD O O O O B-GENEPROD I-GENEPROD O O O O B-CELL_TYPE O O O O O O O B-EXP_ASSAY I-EXP_ASSAY O O B-GENEPROD I-GENEPROD O O O O O O O O O O O O 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-EXP_ASSAY I-EXP_ASSAY O O O O O O O O O O O

D) Quantitative high-resolution mass spectrometry was used to resolve the proteome of Ptpn6/11fl/fl and GzmBcre Ptpn6/11fl/fl CTLs (day 7). The plot depicts protein copy number in control and GzmBcre Ptpn6/11fl/fl cells. Significantly altered proteins are indicated in red; selected examples are annotated. Results are based on n = 3 biological replicates and two-tailed, unequal-variance t-test on log10 transformed copy number per cell values was used to compare differences between experimental groups.


G ) Graph illustrates of DAPI + dead of Ptpn6 / 11fl / fl GzmBcre Ptpn6 / 11fl / fl CTLs ( gated on CD8 + T ) at indicated days as measured by flow cytometry .

O O O O O B-SMALL_MOLECULE O O O B-GENEPROD O O O O B-GENEPROD I-GENEPROD O O O O B-CELL_TYPE O O O B-CELL_TYPE I-CELL_TYPE I-CELL_TYPE O O O O O O O B-EXP_ASSAY I-EXP_ASSAY O

G) Graph illustrates the percentage of DAPI+ dead cells of Ptpn6/11fl/fl and GzmBcre Ptpn6/11fl/fl CTLs (gated on CD8+ T cells) at the indicated days as measured by flow cytometry.


H ) copy number of Tnfrsf1b Mcl - 1 in GzmB cre Ptpn6 / 11fl / fl control CTLs , as estimated through high resolution mass spectrometry .

O O B-EXP_ASSAY I-EXP_ASSAY O B-GENEPROD I-GENEPROD I-GENEPROD I-GENEPROD O B-GENEPROD I-GENEPROD I-GENEPROD O O O O O B-CELL_TYPE O O O O B-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY I-EXP_ASSAY O

H) Protein copy number of Tnfrsf1b and Mcl-1 in GzmB cre Ptpn6/11fl/fl and control CTLs, as estimated through high resolution mass spectrometry.