Skip to main content
. Author manuscript; available in PMC: 2009 Jun 22.
Published in final edited form as: FEBS Lett. 2007 Jul 23;581(22):4153–4158. doi: 10.1016/j.febslet.2007.07.021

Figure 4. Heregulin-induced chromatin remodeling at the utrophin-A promoter.

Figure 4

Serum starved C2C12 cells were treated with 2nM heregulin at time points indicated and analyzed by western blots (A) using phosphohistone H3(Ser10) antibody reveals increased phosphorylation. Anti-H3 antibody was used as control for equal loading. The ratio of band intensities of phosphohistone and corresponding histone showed extent of phosphorylation (B). Immunofluorescence (C) using these antibodies demonstrated increases in global phosphorylation of histone H3 and increased punctate, speckling indicative of activation of promoters. Scale bar = 25μm. ChIP (D) was performed on serum starved C2C12 cells that were transfected with MSK1 followed by incubation with 2nM heregulin. Lysates were analyzed using no antibody (top lane) or phosphohistone H3(Ser10) antibody (middle lane). Radioactive PCR using primers from the utrophin-A promoter region revealed that heregulin-induces chromatin remodeling at the utrophin-A promoter. Aliquots of inputs were used as control (lower lane). The band intensities were quantified and the ratio of phosphohistone and corresponding input band as the measure of occupancy of phosphohistone H3 at utrophin promoter showed ~1.4 fold increase upon heregulin treatment (3E).