FIG. 5.
ACTR is selectively recruited to the promoters of cell cycle genes. ChIP analysis of ACTR occupancy on E2F-responsive genes in T-47D cells infected with adeno-ACTR-HA (labeled with an A) and adeno-GFP (labeled with a G), or treated with 10−7M E2, after cells were hormone deprived and treated with ICI. Cells were harvested 30 and 42 h after infection. The ChIP assay was performed using α-HA (A) or α-ACTR (B) antibodies. (C) Fractions (5%) of total soluble chromatin preparations made from cells treated for panels A and B were taken before immunoprecipitation and used as input; the DNA was isolated in the same way as for the ChIP assay. Usually 1/10 of the DNA isolated from the input and the ChIP samples was used as template for generation of the PCR products presented. Regions of genomic sequence analyzed by PCR are indicated on the schematic of each gene promoter.