Fig. 4: Insulin and HDAC inhibition promote transition of PD1^hi Tph cells to IL7R⁺ memory T cells.

A Transition model of PD1^hi Tph cells to the IL7R⁺ T memory cells, influenced by insulin and HDAC inhibition. B Box plot of protein cytokine levels in supernatants of CD4⁺ cells (n = 12) stimulated with insulin (25 nM) and HDACi (50 µg/mL) for 24 h, by ELISA. Boxes indicate IQR and whiskers indicate the minimum and maximum values. P-values are calculated by Wilcoxon Sign rank test. C Gating strategy to CD4⁺ T cell subset analysis by flow cytometry. D Box plot of PD1 and CD27 expression in CD4⁺ cells (n = 8) stimulated by insulin and HDACi, by mean fluorescence intensity (MFI). Boxes indicate IQR and whiskers indicate the minimum and maximum values. P-values are calculated by Wilcoxon Sign rank paired test. E Heatmap of expression difference of IL7R-related signaling molecules in CD4⁺ cells after in vitro stimulation with insulin (25 nM), HDACi (50 µg/mL), IFNγ (50 ng/ml), and survivin inhibitor YM155 (10 μM), by RNA-seq. Genes connected to cis-RE containing survivin-H3K27ac co-deposition are indicated bold. Asterisks indicate RNA-seq p-values * < 0.05, ** < 0.01, *** <0.001, **** <0.0001. F Heatmap showing expression difference, by log2FC, of cluster markers of Tph cluster after insulin stimulation, HDAC inhibition. In bold are the genes supervised by a cis-RE containing survivin-H3K27ac deposition. P-values are calculated by DESeq2 analysis. Asterisks indicate nominal p-values * < 0.05, ** < 0.01, *** <0.001, **** <0.0001.