Figure 2. AAV2 induced apoptosis/cell death of MDA-MB-435 cells correlates with (A) Rep protein expression. For detecting Rep proteins in immunoblots, total protein extracts were prepared and 60 μg of total protein extracts from AAV2-infected and mock MDA-MB-435 breast cancer cells were resolved in a 7.5% SDS-PAGE gel and detected with an AAV2 Rep specific antibody. Results are representative of three individual experiments. t, time; +, AAV2-infected; −, mock. Actin is used as a loading control. (B) AAV2 genome replicates in virus infected MDA-MB-435 cells. Southern blot analysis to detect 4.7 kb AAV2 replicative form monomer representing active genome replication in the virus infected breast cancer cell line. A total of 5 μg of total DNA was then detected with AAV2 genomic DNA as probe. One hundred nanograms of total DNA isolated from cells was used as loading control. Results shown are representative of 3 individual experiments. t, time; +, AAV2-infected; −, mock. *Low levels of AAV2 genome replication depicted by the 4.7 kb band on days 6 and 13.