Figure 1.

BNIP3 overexpression induces H9c2 cardiomyoblast cell apoptosis. H9c2 cells were transfected with differing amounts (2, 4, 6 μg) of BNIP3-containing plasmids for 24 h. Cells were harvested for different assays. (A) Western blot showed that BNIP3 protein expression increased in a dose-dependent manner; (B) The degree of DNA fragmentation increased in proportion to the level of B-cell lymphoma 2 (BCL2)/adenovirus E1B 19 kDa (BNIP3) expression; (C) JC-1 stain presented mitochondria integrity. The merging of normal (red) and damaged (green) mitochondria represented the integrity ratio. We found that there was a significant fluorescence emission shift from red to green in cells that overexpressed BNIP3, indicating that BNIP3 overexpression led to mitochondrial damage and DNA fragmentation and, hence, activation of the apoptotic pathway.