Prevention of ALP-induced cytochrome c release and apoptosis by overexpression of Bcl-XL. (A) Western blot analysis for Bcl-XL. Cell lysates from Neo- (MM144-Neo) and Bcl-XL–(MM144-Bcl-XL) transfected MM144 cells were subjected to SDS-PAGE and immunoblotted with a Bcl-XL specific antibody to analyze Bcl-XL content. β-Actin was used as a loading control. Experiments shown are representative of 3 performed. (B) Neo- and Bcl-XL–transfected MM144 cells were incubated for 24 hours with 10 μM edelfosine or perifosine, and apoptosis was then quantitated as the percentage of cells in the sub-G1 region by flow cytometry. Untreated control cells were run in parallel. Data shown are means ± SD of 4 independent determinations. (C) Western blot analysis of cytochrome c in cytosolic (CYT) and mitochondrial (MIT) extracts from MM144-Neo and MM144-Bcl-XL cells untreated (−) or treated (+) for 15 hours with 10 μM edelfosine or perifosine. Cytochrome oxidase subunit II (Cyt ox) was also analyzed in the mitochondrial extracts as a control for mitochondrial protein loading. Experiments shown are representative of 3 performed.
