Extended Data Fig. 6: α-KG supplementation restores Treg function and aTreg-associated functional gene expression in TKT deficient Tregs.
a, Flow cytometric analysis (left) and quantification (right) of in vitro suppressive assay of Tregs isolated from 7-week-old WT mice. Tregs were pretreated with DMαKG or vehicle for 4 h and co-cultured with CTV-labelled Tconvs for 3 days. n=4 cultures. b, RT-PCR analysis of indicated genes in WT Tregs, cKO Tregs and DMαKG-treated WT and cKO Tregs. n=3 cultures. c, Global 5hmC/dG levels in Tregs treated with or without DMαKG in vitro. n=3 cultures. d, Representative flow cytometric analysis of CD62L and CD44 expression in CD8+ T cells in the spleen of 6-week-old WT and cKO mice supplemented with or without DMαKG in drinking water. n=7 mice. e, Global 5hmC/dG levels in Tregs isolated from WT and cKO mice supplemented with or without DMαKG in drinking water. n=3 independent samples. f, Heatmap of RNA sequencing of selected genes differentially expressed between WT and cKO Tregs from mice supplied with or without DMαKG in drinking water. g, RT-PCR analysis of indicated genes in Tregs isolated from WT and cKO mice supplemented with or without DMαKG in drinking water. n=3 independent samples. Data are representative of two (a,b,c,g) independent experiments. Data are shown as mean ± s.d.. P value are determined by two-way ANOVA followed by Sidak’s multiple-comparisons test. ns, not significant.
