Fig. 4

RelA regulates the expression of miR-30a. a Linear correlation between miR-30a and TLR4 expression at 0 h, 24 h, 48 h, and 72 h (n = 3 per time point, compared to 0 h) during Th17 differentiation in vitro. b Quantitative PCR analyses of miR-30a expression in Th17 cells from peripheral blood lymphocytes of wild type (WT) and TLR4^−/− mice. c The relative levels of RelA (Western blot) and miR-30a (quantitative PCR) in Th17 cells from peripheral blood lymphocytes of WT EAE mice at different scores (n = 5 per group, mice at score 0 are used as control for RelA and miR-30a respectively). d The relative levels of RelA (MFI analyses of flow cytometry) and miR-30a (quantitative PCR) in Th17 cells from peripheral blood lymphocytes of healthy volunteers (ctrl) and MS patients (n = 8 per group). e–i CD4⁺ naïve T cells from WT mice are cultured in Th17-polarizing conditions with LPS, TPCA1, or Helenalin for 3 days, then miR-30a expression in the cultured CD4⁺ cells is analyzed by quantitative PCR (e); percentage of stimulated IL-17⁺ cells is tested by flow cytometry (f, g); doses of IL-17A, IL-17F, and GM-CSF in culture supernatants are measured by ELISA (h); and Rorγt mRNA level is analyzed by quantitative PCR (i). CD4⁺ naïve T cells cultured in Th17-polarizing conditions are used as control (ctrl). Data are presented as mean ± standard deviation. *P < 0.05. **P < 0.01. ***P < 0.001; Student’s t test (b, d), one-way ANOVA (c, e, g–i). Data are representative of three experiments done in triplicate