Figure 2.

S. clandestina extract induces apoptosis in MG‐63 cells. Cytosolic nuclear and mitochondrial fractions were separated by SDS‐PAGE and analyzed by western blotting using (A) anti‐PARP‐1, anti‐caspase‐3, ‐7, and ‐9 and (B) anti‐BAX, anti‐Bcl2, anti‐cyt‐c, and anti‐BID antibodies. The purity of fractions was assessed by immunoblotting with anti‐β‐actin, A, or with anti‐porin, B, monoclonal antibodies. (C) Cells treated with and without S. clandestina extract and also pretreated with the cell‐permeable broad‐spectrum caspase inhibitor z‐VAD‐fmk (100 μM); left: quantification of the percentage of apoptotic nuclei was obtained by fluorescent staining with 4′,6‐diamidino‐2‐phenylindole (DAPI) (means ± SD; n = 6); P < 0.001 by one‐way ANOVA; values with shared letters are not significantly different according to Bonferroni/Dunn post hoc tests; right: viable cell number evaluation after 24‐h treatment. Statistically significant differences between cells treated with an aqueous extract plus z‐VAD‐fmk and with S. clandestina extract alone (^* P < 0.01) or with untreated cells (§P > 0.05) by Student's t‐test.