FIGURE 9.

SOUL depolarizes mitochondrial membrane in vivo in H₂O₂-treated NIH3T3 cells. Mock-transfected (pcDNA) and SOUL- and ΔBH3-SOUL-overexpressing cells were co-transfected or not with vectors expressing cyclophilin D siRNA (A) or Bcl-2 protein (B). All cells were exposed or not to 100 μm hydrogen peroxide for 3 h, and then the medium was replaced with a fresh one without any agents and containing 1 μm JC-1 membrane potential-sensitive fluorescent dye. After 10 min of loading, green and red fluorescence images of the same field were acquired using a fluorescent microscope equipped with a digital camera. The images were merged to demonstrate depolarization of mitochondrial membrane potential in vivo indicated by loss of the red component of the merged image. Representative merged images of three independent experiments are presented.