Fig. 7
STAT3 and mTOR mediate Butyrate-induction of Th1 cell expression of Blimp-1 and IL-10. a, b Th1 cells were cultured with anti-CD3 and anti-CD28 mAbs in the presence or absence of C4. The cell lysates were prepared, and Western blot was used to determine p-mTOR (S2448, 15 min), p-SATA3 (Y705, 15 min), and p-ERK1/2 (137F5, 15 min), with β-actin as reference. c–f Th1 cells were transfected with lentiCRISPR plasmids to knock out Stat3, mTor, and Mek1. After 24 h, the expression of relative genes (c) was examined by qRT-PCR. The cells were cultured with anti-CD3 and anti-CD28 mAbs in the presence or absence of C4 (n = 3/group). The expression of Prdm1 (d) and IL-10 (e) was examined at day 2 by qRT-PCR. f T-cell cytokine production was determined at day 5 by FACS. Both representative FACS profile and barchart of IL-10+ and IFNγ+ T-cells were shown. The results are shown as mean ± SD of three samples and are representative from one of three experiments performed. *p < 0.05, **p < 0.01 one-way ANOVA test
