FIG. 6.

M11L specifically inhibits Bak-induced apoptosis. HEKM11L and HEKneo cells were transfected with increasing amounts of the HA-Bak expression plasmid as indicated. Cells were harvested after 24 h posttransfection. (A) Apoptotic cells were counted by annexin V-PI staining as measured by fluorescence-activated cell sorter analysis. (B) Western blot analysis of HA-Bak expression, decrease in full-length Bid levels, and cleavage of caspase-3 and PARP in total-cell extracts (50 μg/lane) at various time points after transient transfection of HEKM11L and HEKneo cells with either a vector encoding HA-Bak or the empty control vector. Actin was used as a loading control. HA-Bak has an NH₂-terminal HA tag, providing an additional molecular mass of approximately 3 kDa relative to that of endogenous Bak.