Fig. 3: DMF enhances PI3K-AKT-T-BET-signaling to diminish IL-17 and RORγt in Tc17 cells.

a GSEA examining the enrichment of genes associated with PI3K-AKT-mTOR-signaling in human CD8⁺CD45RA⁻ T cells upon stable response to DMF therapy based on MSigDBv6.1 (dataset from Fig. 2f). b Flow cytometry of IL-17A in murine Tc17 cells differentiated for 72 h ± DMF, ± 1 µM Ly294002 (fold IL-17A induction by Ly294002, a PI3K inhibitor). c, d Flow cytometry of P-AKT(S473) c and P-AKT(T308) d in murine Tc17 cells differentiated for 48 h ± DMF, DMF + GSH, ± 1 µM AKTi (AKT-1/2 inhibitor) or DMF + AKTi. e Flow cytometry of P-FOXO1(T24)/FOXO3a(T32) in Tc17 cells differentiated for 48 h ± DMF or DMF + GSH. Bars to the right in c, d, and e show fold MFI normalized to the respective control, which was arbitrarily set to 1. f Heatmap of color-coded z-scores from the rlog transformed, batch-corrected FOXO1 target genes according to Michelini et al⁴⁴ in Tc17 cells, (dataset from Fig. 2a). g Flow cytometry for IL-17A and IFN-γ in WT and Tbx21^−/− Tc17 cells differentiated for 72 h with indicated treatment (fold IL-17A induction by AKTi). h Flow cytometry for RORγt in WT and Tbx21^−/− Tc17 cells differentiated for 72 h with indicated treatment. Bars show fold RORγt expression (MFI normalized to respective control, which was arbitrarily set to 1). i Flow cytometry for IL-17A in WT and Tbx21^−/− Tc17 cells differentiated for 72 h (fold IL-17A inhibition by DMF). Bars show mean ± s.d. from six c, five to four b, d, e and g–i combined experiments; individual values are plotted. In b, g, i *p < 0.05, **p < 0.01 evaluated by two-tailed, unpaired t-test, in h *p < 0.05 by two-tailed unpaired t-test with Welch’s correction, in (d and e) *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA followed by Tukey’s HSD multiple comparison test, in c *p < 0.05, **p < 0.01, ***p < 0.001 by one-way Welch’s ANOVA with Games-Howell multiple comparison test.