(A) ATR-101 has distinct effects in combination with different inhibitors of electron transport versus ATP synthase on cellular ATP levels. The ATP levels and SYTOX fluorescence of H295R cells that were cultured with the indicated concentrations of H295R cells were cultured with the indicated concentrations of rotenone, antimycin A, sodium azide, CCCP and oligomycin (from top to bottom) in combination with different concentrations of ATR-101 for 20 hours are plotted.
(B) ATR-101 has distinct effects in combination with uncouplers versus inhibitors of F1F0-ATPase on the mitochondrial membrane potential. The JC-1 fluorescence of H295R cells that were cultured with 20 μM ATR-101 and 10 μM CCCP, 100 ng/ml oligomycin or 3 μM DCCD (from top to bottom) separately and in combination for 3 hours are plotted. The graphs show the means and standard deviations of three replicate cultures and are representative of two independent experiments (*: p<0.05 vs vehicle, #: p<0.05 vs ATR-101; two-tailed unpaired Student’s t test).