Figure 7
Pristimerin induces apoptosis in imatinib-sensitive and -resistant CML cells by eliciting mitochondrial damage. (A, B, C) CML cells were exposed to pristimerin with concentrations ranging from 0 to 600 nM for 24 hours (A), or 600 nM pristimerin for various durations (B), then the cells underwent Annexin V/PI double staining for apoptotic cell death assay. Parallel samples were lysed for immunoblotting analysis with antibody against caspase-3 and PARP (C). (D) Pristimerin induced mitochondrial release of cytochrome c and apoptosis-inducing factor (AIF). KBM5, KBM5-T315I or K562 cells were treated with pristimerin at 600 nM, 600 nM and 800 nM, respectively, for 0-24 hours; levels of cytochrome c in the cytosolic extracts prepared using a digitonin buffer were detected by Western blot analysis. (E) Western blot analysis of apoptosis-related proteins such as Bcl-2, Bcl-XL, Mcl-1 and survivin was shown. The indicated CML cells were exposed to pristimerin for 24 h. (F) Immunoblotting analysis of apoptosis-related proteins in response to pristimerin (600 nM) for different durations was shown.