2.6. Western blot analysis

PC3 cells treated with various crude extracts were lysed at the time indicated. Equal amount of cell lysates were electrophoresed on SDS-polyacrylamide gels. The proteins were transferred to the PVDF membrane and incubated with primary antibodies recognizing caspase-3, caspase-9, PARP, p21, p53, and β-actin. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibodies for 60 minutes and developed using enhanced chemiluminescence (ECL; Millipore, Watford, Herts, UK) [14].