Figure 5.

ChIP assays detect ERα and P-TEFb components in the attenuation region of the MYB gene. (A,B) ERα binding to the MYB intron-I attenuation region and the pS2 promoter. In total, 0.25, 0.5 or 1.0 µg of anti-ERα antibody, no antibody (mock) or control IgG (1.0 µg) was added to MCF-7 cell extracts and ChIP assays were performed as described in ‘Materials and Methods’ section. (C) P-TEFb complex components are selectively recruited to the MYB intron-1 region. Anti-CyclinT1 and anti-CDK9 antibody (0.25 or 0.5 µg) or 0.5 µg of IgG was added to the MCF-7 extract for ChIP assays. The amount of DNA bound to ERα, CyclinT1 and CDK9 was detected by qPCR using MYB intron- and pS2 promoter-specific primers and the results are plotted as percentage of input.