Analysis of cellular viability in the presence of respiratory inhibitors was performed using MTS assay (Promega, G3580) according to the manufacturer's instructions. Briefly, cells were seeded at a density of 2 × 104 cells per well in Matrigel-treated 96-well culture microplates in 100 μl of culture media and incubated for 4 h at 37 °C. Next, the mitochondrial respiratory inhibitors FCCP or 0.5 μM Antimycin A+0.5 μM Rotenone (Sigma) were added at the indicated concentrations. After 24 h of compound/DMSO treatment, 20 μl of MTS reagent was applied to each well of 96-well plate. Absorbance at 490 nm was recorded after 2 h incubation. Sextuplets were prepared for each condition.
