Fig. 7. Targeting of proAR to the nuclear envelope causes heterochromatin assembly. (A) HeLa cells transiently expressing wild-type AR were incubated with TPA and treated with Triton X-100, DNase I and/or NaCl. Cells were then stained with anti-AR-N pAb and anti-lamin A/C mAb. Arrowheads indicate AR-expressing cells. (B) HeLa cells transiently expressing wild-type AR were incubated with TPA and then stained with anti-AR-N pAb, anti-dimethylated H3K9 (H3K9Me2) mAb, anti-trimethylated H3K9 (H3K9Me3) pAb and anti-HP1b pAb. Cells were counterstained with Hoechst 33342. Each panel shows representative cells that are untransfected (top row) and transfected (bottom row). (C) Cell lysates of HeLa cells transiently expressing wild-type AR or AR-C11 treated with or without TPA were analyzed by western blotting with anti-AR-N pAb, anti-H3K9Me3 mAb and anti-β-actin mAb. Scale bars: 5 µm.