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. 2014 Jun 3;289(30):20502–20515. doi: 10.1074/jbc.M114.569889

FIGURE 6.

FIGURE 6.

a, nontargeting shRNA (NT) and TBL1XR1 targeting shRNA RS4;11 were treated with 500 μg/ml of prednisolone for 3 h. Chromatin-enriched fractions were extracted followed by immunoblotting to analyze the levels of chromatin-bound TBL1XR1, GR, NCoR1, and HDAC3 proteins. Heterochromatin protein HP1 was used as a loading control for the chromatin-bound fraction, tubulin was used as a control for the cytosolic fraction, and PARP1 was used as a control for the soluble nuclear fraction. 2.5% of 4 million cells (100,000 cells represented on the gel) was loaded for the whole cell lysate, whereas 50% of the chromatin-enriched fraction of 4 million total cells (2 million cells represented on the gel) was loaded for the chromatin fractions. Each Western blot is from one representative experiment. The experiment was repeated at least twice in each cell line with similar results. b, Western blot for the GR in nontargeting shRNA (NT) and TBL1XR1 knockdown RS4;11 lines.