Extended Data Fig. 4: PD-1 controls ACLY-dependent epigenetic reprogramming.

a, H3K27ac in CD4 + GFP + T cells of ITK-SYK^CD4-creERT2 or C57BL/6 mice on d5 after TAM +/− anti-PD-L1 antibody treatment. b, c, Non-labeled (b) and labeled (c) acetyl-CoA determined by targeted LC-MS/MS in ITK-SYK⁺ T cells +/− PD-1 cultured with uniformly labeled [U-¹³C]glucose (11 mM) for 4 h in vitro (n = 4 mice genotype). Normalization to viable cell numbers. P=two-sided Student’s t-test; mean ± SEM. d, [U-¹³C]glucose-derived ¹³C-acetyl groups on histones determined by orbitrap LC-MS/MS in ITK-SYK⁺ T cells +/− PD-1 cultured with uniformly labeled [U-¹³C]glucose (11 mM) for 4 h in vitro (n = 3 mice genotype). Normalization as in (b, c). P=two-sided Student’s t-test; mean ± SD. e, H3K27ac in ITK-SYK⁺ T cells +/− PD-1 after 3 h in vitro incubation with BMS-303141 (‘iACLY’, 15 μM) +/− acetate (100 μM; n = 3 biological replicates). P=paired two-sided Student’s t-test; mean ± SD. f, Viable ITK-SYK⁺PD-1^- cells after 72 h in vitro culture with BMS-303141 (‘iACLY,’ 5 μM) or DMSO (n = 3 biological replicates). P=paired two-sided Student’s t-test. g, Viable ITK-SYK⁺ cells in the blood of sgACLY or sgMOCK-injected mice (n = 4 per group). P=two-sided Student’s t-test. h, Viable ITK-SYK⁺ cells +/− PD-1 after 3-day in vitro incubation with FT113 (1 μM) or DMSO (n = 3 biological replicates). P=two-sided Student’s t-test; mean ± SD. i, Viable ITK-SYK⁺ cells in presence of NEO2734 (50 nM). Experiment as in (e). P=two-sided Student’s t-test; mean ± SD. j, Percentage of H3K27ac ChIP-seq peak distribution over different genomic features as determined from the experiment in (d). k, Genome-wide average H3K27ac scores in a 3 Kb window around the TSS. ChIP-seq was performed as indicated in Fig. 4h. ChIP-seq, chromatin immunoprecipitation sequencing; TSS, transcription start site. a, Representative data from two independent experiments with n = 3 biological replicates per genotype. b, c, one experiment, n = 4 biological replicates. d, Data from one experiment. e, Representative data from one experiment with n = 3 biological replicates per genotype. f, Representative data from two independent experiments. g, Data from a single experiment. h,i, Representative data from three independent experiments with n = 3 biological replicates per genotype. j,k, Data from one experiment.

Source data