Effect of CD437 on the release of cytochrome c into the cytosol and the mitochondrial transmembrane potential of NB4 and NB4.437r cells. (A) The amount of cytochrome c released in the cytosol of NB4 and NB4.437r cells (5 × 105/mL) after challenge with vehicle (control) or CD437 (10−6mol/L) for the indicated amount of time was measured by Western blot analysis using a specific polyclonal antibody. The proportion of apoptotic cells was determined by DAPI staining on an aliquot of the cells used for the cytochrome c release assay. (B) NB4 or NB4.437r cells were preloaded with the fluorescent dye 3,3′-dihexiloxadicarbocyanine iodide (DiOC6), and treated with vehicle,2 CD437 (10−6mol/L),3 the proton translocator carbonyl cyanide p-(trifluorometoxy)phenylhydrazone (250 nM) FCCP,4 or CD437 + FCCP5 for 30 minutes. The transmembrane potential was assessed by flow cytometry. The fluorescence level in the absence of DiOC6 is indicated by tracing.1 Data are representative of at least 2 independent experiments with similar results.
