Figure 3

Regulation of IL-10 gene expression by MEF2D. (A) LPS-induced time-dependent changes of MEF2D protein and cytokines in BV2 cells. BV2 cells treated with LPS as described in Figure 2A were analyzed by immunoblotting for MEF2D and by qPCR for TNF-α and IL-10 mRNA levels. Data from three independent experiments were expressed as the mean ± SEM and analyzed by one-way ANOVA (*P < 0.05; **P < 0.01). (B) Identification of a conserved MEF2 binding site in the IL-10 gene promoter of different species. The underlined sequence indicates the MEF2 binding site. Black-shaded areas show the conversed MEF2 binding site in the IL-10 gene promoter of different species. (C, D) Binding of MEF2D to the IL-10 gene promoter region containing the conserved site in BV2 cells. BV2 cells treated with or without LPS for 24 h were analyzed by ChIP. The result was showed by standard endpoint PCR (C) and qPCR (D). Data from three independent experiments were expressed as the mean ± SEM and analyzed by one-way ANOVA (*P < 0.05; **P < 0.01).