Figure 7. Inhibition of pERK or pAkt does not affect mitochondrial membrane potential. JC-1 analysis of HLE-B3 cells treated with LY294002 or UO126. Cells were initially seeded onto 35 mm dishes and allowed to grow to semiconfluence. The cells were then pretreated with 25 µM LY294002 or 10 µM UO126 in 0.05% DMSO for 3 h in hypoxia to determine the state of mitochondrial membrane potential. Control cells were treated with 0.05% DMSO in serum-free media. After hypoxic exposure, the media were replaced with fresh, oxygenated serum-free media containing 5 µg/ml JC-1 for 30 min in atmospheric oxygen. The media were removed, and fresh serum-free media without inhibitor were added to the cells. Serial confocal imaging indicated no significant difference in the green/red ratio between the cells treated with LY294002 (A) or UO126 (B) and control cells (Student t test, p>0.05).