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. 2018 Oct 10;10:4421–4438. doi: 10.2147/CMAR.S176088

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© 2018 Chen et al. This work is published and licensed by Dove Medical Press Limited

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The effect of LINE-1 ORF-1p on PXR cytoplasmic/nuclear translocation and recruitment to the promoter or enhancer region of CYP3A4.

Notes: MHCC97-H cells were transfected with an empty vector, the expression vector for LINE-1 ORF-1p (A and C), control siRNA or the siRNA for LINE-1 ORF-1p (B and D) and treated with or without 10 µmol/L rifampicin. (A and B) Cells were fractionated into cytoplasmic fractions and nuclear fractions. The fractions were probed using anti-LINE-1 ORF-1p and anti-PXR antibodies. Lamin A/C was chosen as the nuclear marker and ß-actin as the cytoplasmic marker. (C and D) Cells were harvested for ChIP experiments. Immunoprecipitated DNA was PCR amplified using primers that were consistent with the PXR binding sites in the CYP3A4 promoter or enhancer region.

Abbreviations: ChIP, chromatin immunoprecipitation; PXR, pregnenolone X receptor.