Fig. 4

FTY720 downregulates Akt and GSK3β phosphorylation and induces epigenetic modification. a Western blot analysis of cell extracts from FTY720-treated (T-FTY720) and control T (T-CTL) cells at the indicated time points and analyzed for phosphorylated Akt (p-Akt) and total Akt (tot-Akt). β-actin served as a loading control. A representative image from one experiment is shown. b Immunoblot of phosphorylated GSK3β (pGSK3β) and total GSK3β (tot-GSK3β) in cell extracts from T-FTY720 and T-CTL cells at the indicated time points. β-actin served as a loading control. A representative image from one experiment is shown. c Immunoblot of β-catenin in cell extracts from FTY720 and T-CTL cells at the indicated time points. β-actin served as a loading control. A representative image from one experiment is shown. d qPCR analysis of chromatin-immunoprecipitated T cells showing methylation status of H3K9 and H3K27 on the IFNG promoter region (n = 7). Data are shown as mean ± SEM. *p < 0.05, paired non-parametric t tests. e qPCR analysis of chromatin-immunoprecipitated T cells showing the methylation status of H3K9 and H3K27 on the GZMB promoter region (n = 7). Data are shown as mean ± SEM. *p < 0.05, paired non-parametric t tests