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Substrates were added in a stepwise fashion in the following order (final concentrations) 0.2 mM palmitoyl carnitine, 1 mM malate, 4 mM ADP, 0.4 mM octanoyl carnitine, 5 mM pyruvate, 10 mM glutamate, 10 mM succinate, 1 µM CCCP, 2 µM rotenone, and 5 µM Antimycin A