Fig. 3: Restoration of HIF1α and intracellular acetyl-CoA rescues IFN-γ production in Hif1α^–/– T cells.

A, B Protein expression of HIF1α in WT and Hif1α^{− / −} CD4⁺ T cells successfully transduced (GFP⁺ ) with empty retroviruses (EV) or retroviruses expressing WT or triple-mutant Hif1α (TM) under hypoxia (A). GFP⁺ T cells from (A) were activated under hypoxia and analyzed for IFN-γ production (B) (N = 3 per group, ****p < 0.0001)). C Intracellular acetyl-CoA in activated WT and Hif1α^{− / −} CD4⁺ T cells (N = 6 per group, ****p < 0.0001). D–E Intracellular acetyl-CoA (D) (N = 6 per group, *p = 0.022) and IFN-γ production (E) (N = 5 per group, **p = 0.0081) by activated WT and Hif1α^{− / −} CD4⁺ T cells, cultured with or without 20 mM sodium acetate (NaAc) added on Day 2 post-activation. F Cell death of MB49 cells cultured with activated WT and Hif1α^{− / −} CD4⁺ T cells at the ratio of 1:2 for 48 h was analyzed by 7-AAD/Annexin V staining (N = 12 per group, ****p < 0.0001). G Cell death of MB49 cells co-cultured with activated WT and Hif1α^{− / −} CD4⁺ T cells pretreated with or without NaAc for 48 h was analyzed by 7-AAD/Annexin V staining (N = 4 per group, ****p < 0.0001). H,I H3K9Ac enrichment at Ifng promoter and upstream regions (H) and Ifng mRNA expression (I) (N = 3 per group, *p = 0.0123) in activated WT and Hif1α^{− / −} CD4⁺ T cells treated with solvent (UnTx) or 20 mM of NaAc. A two-sided Student’s t-test was used in (C and F) for statistical analyzes. Two-way ANOVA with Šídák’s multiple comparisons test (with adjustment) was used for (B, D–E, G) and I. All experiments were repeated at least twice. Pooled results shown in the dot plots and bar graphs depicted means ± SEM for all the samples in each group, with each dot denoting an independent sample. Source data were provided in the Source Data file.