Fig. 4.

Oleanolic acid 3-acetate induces the release of cytochrome c and AIF from the mitochondria with a concomitant activation of caspase-3,caspase-7,caspase-8, and PARP in SKOV3 and HEC-1A cells. SKOV3 and HEC-1A cells were exposed to the indicated concentration of oleanolic acid 3-acetate for 24 h. (A) Total cell lysates were subjected to Western blot analysis with the specific antibody against PARP, cleaved PARP, cleaved capase-7, cleaved caspase-8, cleaved caspase-3, Bcl-2, and β-actin. β-Actin was used as a loading control. (B) The (C) cytosolic fractions and (D) mitochondrial fractions were separated and subjected to Western blot analysis for AIF, cytochrome c, HSP60, and β-actin. The bars represent the results of densitometric analysis of band intensities in (B) corresponding to mitochondrial and cytosolic cytochrome c by using MultiGuage software. Data were normalized to control and presented as means ± SD of at least triplicate determinations. *p < 0.05 versus control (without oleanolic acid 3-acetate treatment). AIF, apoptosis-inducing factor; PARP, poly(ADP-ribose) polymerase; SD, standard deviation.