Figure 2.

MtDNA depletion decreases BRCA2 protein expression. (a) Wild-type and mtDNA-depleted cells were assessed for BRCA2 and BRCA1 protein expression by western blotting. (b) Total DNA was extracted from LNCaP and C4-2 prostate carcinoma cells, and mtDNA was amplified by long PCR (left panel) or quantified by real-time PCR (right bar graph) using primers for D-loop or Cox II. At the top right panel, BRCA2 levels were analyzed by western blotting. (c) MtDNA-depleted cells were transiently transfected with BRCA2 cDNA or empty vector (control), treated with AG014699 for 48 h, and Rad51 foci (left panel) and cell survival (right panel) were measured by immunofluorescence and clonogenic assays, respectively. Data are expressed as the percentage of untreated controls. Representative blots of BRCA2 protein levels in transiently transfected cells are shown.