Fig. 9.

Recruitment of FXRα1 and FXRα2 to IR1a and IR1b in intact cells. ChIP assays were carried out using chromatins prepared from Huh 7 cells cotransfected with (A) FXRα1 or (B) FXRα2 with human BSEP promoter reporter wt, IR1a, IR1b, or IR1ab double mutant. After immunoprecipitated with anti-FXR Abs or IgG as negative control, recruitment of FXRα1 or FXRα2 to IR1a or IR1b was detected by PCR using a set of primers, BSEP(-260b) and pGL-R(122), flanking the two IR1 sites in the reporter plasmid DNA. A negative control primer set, BSEP(-995b) and BSEP(-805b), was used to amplify a fragment approximately 700b upstream the IR1 sites. Input chromatin DNA was included as a positive PCR control.