FIGURE 3.

Effect of BNIP3 loss-of-function on mitochondrial mass and respiration in WT and KO MEFs. A and B, quantitative real-time RT-PCR (A) and immunoblot analysis (B) showed down-regulation of BNIP3 mRNA and protein, respectively, by short hairpin RNAs sh80 and sh82 in cells incubated at 20 or 1% O₂ for 24 h (A) or 48 h (B). β-Actin blot showed equal protein loading. NS, nonspecific band. C, D, E, and F, mitochondrial DNA content (C), mitochondrial mass (D), O₂ consumption (E), and ATP levels (F) were measured in MEF subclones that were stably transfected with EV or vector encoding sh80 or sh82 and cultured at 20 or 1% O₂ for 48 h. Data are presented as the mean (±S.E.). ^*, p < 0.05 by Student's t test compared with WT-EV MEFs at 20% O₂;#, p < 0.05 compared with WT-EV MEFs at 1% O₂.