Fig. 1.

BSEP mRNA expression was induced by oxysterol 22(R)-OHC. Human primary hepatocytes derived from three individual donors were treated with 0.1% ethanol (ETOH), 10 μM 22(R)-OHC, 0.1% DMSO, 10 μM CDCA, and rifampicin (RIF) for 30 h. Total cellular RNAs were isolated and subjected to RT-PCR using BSEP-specific primers. The PCR cycles were 30 for donors 1 and 3 and 35 for donor 2. Detection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was included as an internal control. The PCR-amplified products were resolved by agarose gel electrophoresis (1.8%), detected by a Typhoon 9410 Image Scanner, and quantitated by KODAK 1D Image Analysis Software.