Effects of TFAM knockdown using siRNA. A. Mitochondrial membrane potential (Mito Tracker Red) and expression of ATP5A. ATP5A which documents the existence of ATP production, a major mitochondrial function, was detected. In Dex(-)/normoxia (normoxia) the mitochondrial membrane potential was preserved, and ATP5A expression was also found. By TFAM knockdown the mitochondrial membrane potential was no longer preserved, and ATP5A expression was attenuated. Each five independent experiments were carried out. (Scale bar: 20 µm) B. Western blotting of TFAM siRNA-transfected cells. In WB TFAM (29kDa) expression was significantly inhibited by TFAM siRNA. Each three independent experiments were carried out. C. JC-1 staining. On JC-1 staining, the mitochondrial membrane potential decreased by the osteocyte apoptosis induced by TFAM knockdown. Each three independent experiments were carried out. Viable cells: red, apoptotic cells: green. (Scale bar: 200 µm).