Figure 5.

Inhibiting IL-6R abolishes the promoting effect of miR-124 on Th1 polarization. (A) The protein expression of IL-6R in colon tissues after miR-124 agomir or LNA administration in DSS mice, as mentioned in Figures 1, 2, was detected by Western blot (N = 3). Corresponding quantification is also shown. (B–E) Human lymphocytes were isolated and treated with IL-2 (80 ng/mL), anti-CD3e (50 ng/mL), and anti-CD28 (50 ng/mL) for 24 h and then transfected with miR-124 LNA or corresponding scramble control (200 ng/mL) for 60 h. They were then treated with or without IL-6R siRNA (100 nM) for 36 h. The protein expression of IL-6R and its corresponding quantification was analyzed by Western blot (N = 3) (B). IFN-γ and IL-4 expression gated on CD4+ T cells were analyzed by FCM (N = 6 per group), and the IFN-γ/IL-4 ratio and representative flow images are shown in (C,D). Representative Western blots (N = 3) of T-bet and GATA3 and corresponding quantification are shown in (E). Data represent mean ± SEM and were analyzed by one-way ANOVA followed by post-hoc Tukey's test. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001. NS, no significant.