FIGURE 5.

Knockdown (KD) of cyclophilin D sensitizes cells to apoptosis induced with staurosporine or serum starvation. A, stable empty vector-(Puro (for puromycin)) or CypD shRNA-transfected cells were treated with Sts at 2 μm for 6 h or serum starvation (Starv) for 48 h or TNF at 10 ng/ml for 24 h. Cytosolic fractions or whole cell lysates were prepared and normalized for total protein. Cytosolic fractions were subjected to a CytC ELISA assay (A), and cell lysates were subjected to a caspase-3 activity assay (B), as described under “Experimental Procedures.” Data are means ± S.D. (n = 3). * indicates p < 0.05 when compared with puromycin (Puro) controls. UT, untransfected.