FIGURE 7.

The interaction between ACTN4 and GR only occurred in the nucleus in HPCs. A, HPCs were treated with 100 nm dexamethasone for 4 h, and nuclear (N) and cytoplasmic (C) fractions were prepared. Coimmunoprecipitation (IP) was carried out using anti-GR antibodies, followed by Western blotting with the indicated antibodies. B, HeLa cells were transiently transfected with an HA-GR expression plasmid. Nuclear (4) and cytoplasmic (4) lysates were incubated with immobilized GST or GST-ACTN4. Pulldown assays were performed with nuclear extracts expressing HA-GR (lanes 1–3) and with cytoplasmic extracts (lanes 4–6). Pulldown fractions were analyzed by Western blotting with anti-HA antibodies. The expressions of GST fusion proteins were visualized by Coomassie Blue staining.