Figure 4.

Analyses of glucocorticoid and BRD4 actions to Foxp1 expression in bone-marrow mesenchymal stem cells. BRD4 knockdown attenuated glucocorticoid-mediated upregulation of H3K9ac enrichment (a) and BRD4 occupation (b) at Foxp1 promoter along with significant reductions in Foxp1 mRNA expression (c) and protein levels (d). Glucocorticoid-treated cells showed strong Foxp1 immunofluorescence in the nuclear compartment, whereas the immunoreaction was attenuated in BRD4 RNAi-transfected cells (e); scale bar, 8 μm. H3K9ac and BRD4 enrichment in the promoter was probed using ChIP-PCR; Foxp1 expression was quantified using qRT-PCR, immunoblotting, and immunofluorescence staining. Data are expressed as mean ± SEM calculated from 3 experiments. Asterisks * indicate significant differences from the scramble group and hashtag # indicates significant differences from the GC group (p < 0.05). GC: glucocorticoid.