A: Ligand-activated GR releases INHAT components Set/TAF-Iβ and pp32 from the MMTV GREs in HCT116/MMTV cells (left panels) and TAT GREs in HTC cells (right panels)
HCT116/MMTV cells transfected with pRShGRα or the control plasmid and HTC cells were treated with 10−6 M of dexamethasone (Dex). They were fixed with 1% formaldehyde and ChIP assays were performed by using anti-Set/TAF-Iβ, anti-pp32, anti -GRα or control antibodies.
B: Ligand-activated GR releases the wild type Set/TAF-Iβ but not the GR-binding domain-defective Set/TAF-Iβ-(Δ181–225) from the MMTV-GREs in HCT116/MMTV cells.
HCT116/MMTV cells were transfected with His-Set/TAF-Iβ-expressing plasmids and/or pRShGRα. The cells were treated with 10−6 M of dexamethasone (Dex), fixed with 1% formaldehyde and ChIP assays were performed by using anti-His, anti-GRαor control antibodies.
C: Overexpression of Set/TAF-Iβ-(181–225) attenuates GR-induced release of Set/TAF-Iβ from GREs in HCT116/MMTV cells. HCT116/MMTV cells were transfected with 0.5 μg/well of His-Set/TAF-Iβ-expressing plasmid (as “1”), pRShGRα and the indicated amounts (ratios) of the Set/TAF-Iβ-(Δ181–225)-expressing plasmid. The cells were treated with 10−6 M of dexamethasone (Dex), fixed with 1% formaldehyde and ChIP assays were performed using anti-His, anti-GRαor control antibodies.