Research Paper Volume 5, Issue 1 pp 3—17

Figure 2. Casodex-induced TIF response is independent of p53 and AR mutation status. (A, B), Casodex-induced TIF response is independent of p53. LNCaP cells were infected with a dominant-negative p53 (GSE-22) or control retroviral expression vector, and selected for 2-3 d. Cells were then treated (A) with or without 100 μM Casodex for 24 hr, and then analyzed for 53BP1 foci [44], or (B) with or without 10 μg/ml etoposide for 24 hr, then fixed and stained with p21 antibody. (C-E), Casodex induces a TIF response in AR-positive LAPC4 cells with wild-type AR. (C), LAPC4 cells were treated with or without 100 μM Casodex for 24 hr and then co-immunostained with antibodies to γH2AX (green) and TIN2 (red). Colocalization of γH2AX and TIN2 is shown in the ‘merge’ panel. (D), Higher magnification images of a representative Casodex-treated LAPC4 cell stained with antibodies to γH2AX and TIN2. (E), LAPC4 cells and LNCaP cells were treated with and without 100 μM Casodex for 24 hr, then evaluated for the percentage of cells with >10 53BP1 foci/cell. Bars represent the mean ± standard deviation.