Cell proliferation, apoptosis, cell cycle analysis and protein analysis.

<p>(A) Cell viability examined by trypan blue exclusion in LNCaP (LN), LNb4 and PC-3 cells. (B) Determination of apoptosis by flow cytometry. After treatment of cells with Doc, the extent of apoptosis was assessed by annexin V/7AAD assay. (C) MTS assay in PC-3 cells transfected as indicated in figure and treated 48 hours with 5 nM Doc. (D) Graphs from flow cytometry in cells mentioned in (C) demonstrating cycling cell population stained with propidium iodide. Cells were treated with Doc for 48 h or remained untreated and sorted into different stages of mitosis by flow cytometry. (E) Western blotting analysis to show protein expression in transfected cells mentioned in (C). (F) Analysis of protein-protein interaction determined by immunoprecipitation in the presence or absence of Doc or Pac in LN and LNb4 cells. Cells were immunoprecipitated with anti-AR (441) antibody and membranes were probed with anti-c-jun antibody. For control, 20 µg of the cell lysate was used as input and equal loading was confimed with AR antibody.