Monoclonal ANTI-FLAG® M2 antibody produced in mouse

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	Cells were then treated using 10 μM rifampicin or 1‰ DMSO as control sample for 24 h. After the interval, the cells were harvested, and immunoprecipitation was conducted with the Imprint® Chromatin Immunoprecipitation Kit (Sigma‐Aldrich) according to the manufacturer's protocol.

Protocol tips
Cells were then treated using 10 μM rifampicin or 1‰ DMSO as control sample for 24 h. After the interval, the cells were harvested, and immunoprecipitation was conducted with the Imprint® Chromatin Immunoprecipitation Kit (Sigma‐Aldrich) according to the manufacturer's protocol.

Publication protocol

Steroid receptor coactivator (SRC)‐1 recruitment to the HNF4α responsive elements (DR‐2) and E‐box in the OCT1 gene promoter was determined using a chromatin immunoprecipitation (ChIP) assay in the HepG2 cells, which were transfected with pSG5‐hPXR and pSG5‐FLAG hSRC‐1 (3.2 μg in 25 cm2 flask) expression plasmids after one day of cultivation. Cells were then treated using 10 μM rifampicin or 1‰ DMSO as control sample for 24 h. After the interval, the cells were harvested, and immunoprecipitation was conducted with the Imprint® Chromatin Immunoprecipitation Kit (Sigma‐Aldrich) according to the manufacturer's protocol. Anti‐FLAG M2 antibody (Sigma‐Aldrich), normal mouse IgG antibody and anti‐RNA polymerase II (Pol II) antibody were used for precipitation. Levels of SRC‐1 recruitment to the HNF4α response elements and into the E‐box were analysed by qRT‐PCR with primers and probes specifically designed to amplify indicated regions (Figure 6, upper panel, arrows). Data are presented as a relative binding to control vehicle (DMSO)‐treated cell samples (100%) immunoprecipitated with the same antibody. Non‐specific immunoprecipitation with mouse IgG antibody was 1% lower than with anti‐FLAG or the anti‐Pol II antibodies.

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