Figure 3.

MiR-124 promotes Th1 polarization in human peripheral blood lymphocytes. Cells were treated with IL-2 (80 ng/mL), anti-CD3e (50 ng/mL), and anti-CD28 (50 ng/mL) for 24 h and transfected with agomir (300 nM) or miR-124 LNA (200 ng/mL) for 36 h. Before FCM detection, PMA/Ionomycin, and BFA/Monensin mixture were added for 6 h. (A,D) Th1/Th2 (IFN-γ/IL-4) ratio was analyzed and representative flow images gated on CD4+ T cells are shown. (B,E) Representative Western blots (N=3) of T-bet and GATA3 and corresponding quantification. (C,F) Th1-related cytokines IFN-γ and IL-2 and Th2-related cytokines IL-4, IL-6, and IL-13 in the supernatants were analyzed by CBA and ELISA. N = 6–9 per group. All data represent mean ± SEM. ^*p < 0.05, ^**p < 0.01 vs. control (or scramble) group by Student's t-test.