Figure 2. Effect of mitochondrial inhibitors on energy-producing pathways of HepG2/C3A cells.

HepG2/C3A cells were washed twice with serum-free inoculating medium (RPMI-1640 with no phenol red or glucose but containing PS, 0.3 mM glutamine) and resuspended in the same medium at a density of 400,000 cells/mL with varying concentrations of either FCCP or rotenone. Control cells were incubated with 0.003% DMSO, the solvent for both FCCP and rotenone. These cell suspensions were immediately inoculated into PM-M1 at 50 µL per well. After an 18-hour incubation at 37°C under 5% CO₂-95% air, 10 µL of Redox Dye Mix MA was added to each well. After 5 hours with the dye, the plates were photographed. After the plates were photographed, glucose was added to each well of the plates to a final concentration of 5 mM to assess cell viability. The plates were then incubated overnight and photographed again.