Fig. 4.

3Cl-AHPC treatment increases occupancy of SHP at the CYP7A1 and CYP8B1 genes. A and B, HepG2 cells were infected with Ad-flag-SHP; 24 h later, cells were treated with 100 nm 3Cl-AHPC or vehicle for 2 h, and occupancy of SHP and other proteins were examined by ChIP assays. Semi-q-PCR was performed to detect occupancy at the CYP7A1 promoter and the coding region of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a negative control. C and D, HepG2 cells were treated with 200 nm 3Cl-AHPC for 2 h. Occupancy at the CYP7A1 and CYP8B1 genes by endogenous SHP, LRH-1, and RNA polymerase II (Pol II) was examined by ChIP assays followed by semi-q-PCR. Reproducible results were observed from multiple independent ChIP assays as indicated. B and D, Band intensities were determined and the values for control samples treated with vehicle were set to 1.