Fig. 2.

TFEB regulates Treg differentiation in vitro. (A) Immunoblot analysis (Left) and quantitative results (Right) of TFEB in Th17 and Treg cells. (B and C) WT and Tcfeb-deficient naïve CD4⁺ T cells were differentiated under Th17 (B) and Treg (C) conditions for 3 d. Representative flow cytometry plots and frequencies of IL-17⁺ T cells (B) and Foxp3⁺ T cells (C). (D) Activated CD4⁺ T cells were transduced with control retrovirus or TFEB-S211A retrovirus and then differentiated under Treg conditions. Representative flow cytometry plots and frequencies of Foxp3⁺ cells were shown. (E) The occupancies of H3K4me3 in the Foxp3 promoter were determined by chromatin immunoprecipitation and reverse transcription–PCR. (F and G) Representative flow cytometry plot (F) and frequencies (G) of CD4⁺ Foxp3⁺ Treg cells in spleen, pLNs, and colonic lamina propria (LP) of 20-wk-old Tcfeb^fl/fl and CD4^CreTcfeb^fl/fl mice (n = 4). ns, no significance; *P < 0.05, **P < 0.01, ***P < 0.001. Data are representative of three (A, D, E) or four (B, C) experiments. Data are means ± SEM and were analyzed by two-tailed, unpaired Student’s t test.