AAV2 infection of nHMECs does not result in activation of caspases of either the intrinsic and extrinsic pathways of apoptosis. nHMEC monolayer cultures were synchronized in G1 as described, followed by infection with AAV2. Cell pellets were collected each day over a 5 day period. Cells were passaged 1:2 on day 2. Detection of caspases and their cleavage/activation was performed by Western blotting. Sixty micrograms of total protein extracts from AAV2 infected and control nHMEC cells were resolved in SDS-polyacrylamide gel electrophoresis (PAGE) gels. To detect caspase-8 proteins were resolved in a 10% SDS-PAGE gel and detected with a mouse monoclonal antibody (Alexis Biochemicals). To detect the 35 kDa pro-caspase form of caspase-3, proteins were resolved in a 10% SDS-PAGE gel and detected with caspase-3 rabbit monoclonal antibody (Cell Signaling Technology). To detect the pro- (116 kDa) form of PARP, proteins were resolved in a 7.5% SDS-PAGE gel and detected with a rabbit monoclonal antibody (Cell Signaling).