Figure 3

Effect of saposin C on expression/activity of caspases and PARP under serum-starvation stress. Cells were cultured routinely up to 60% confluency, washed with PBS, and incubated in their respective serum-free media supplemented with or without saposin C for 48 h. Cell lysates were prepared as described in Materials and Methods and 75 μg of clarified protein samples was subjected to SDS-PAGE under reducing conditions. Western analysis was carried out using monoclonal antibodies against non-cleaved and cleaved caspases-3, -7, and -9 and PARP. For control loading, membranes were probed or reprobed with anti-actin antibody. Each experiment was performed in duplicate, and the assays were repeated three times.