Fig. 8.

CINPA1 treatment does not allow CAR binding to DNA response elements at the CYP2B6 gene promoter in human hepatocytes. (A) PBREM and XREM (dNR3) in the CYP2B6 promoter region. (B–D) Human primary hepatocytes from three separate donors were treated overnight with DMSO, 1 µM CITCO, 5 µM CINPA1, or 10 µM PK11195. Protein complexes were cross-linked and chromatin was immunoprecipitated by using anti-CAR antibody or control IgG. CAR occupancy at two separate CYP2B6 promoter regions (dNR3 and PBREM) was determined by performing quantitative real-time PCR assays. CAR occupancy at a CAR-free intergenic region around +14 kb was also analyzed as a negative control for recruitment. Fold-enrichment normalized to IgG control was plotted. Data represent mean ± S.D. of three experiments. P values (denoted *, **, or ***) were determined by comparing compound treatments with DMSO-treated samples.