Figure 4.

An electrode sensitive to the TPP moiety of MitoDNP-SUM was conditioned in KHE buffer and 10 μm methyltriphenylphosphonium (TPMP) for two weeks prior to the experiment, then washed extensively before use to remove TPMP. Rat skeletal muscle mitochondria (0.3 mg protein per mL) were incubated in KHE plus 0.3 % (w/v) BSA with stirring at 37 °C. A) Two aliquots of 3 μm MitoDNP-SUM were added to the chamber, and mitochondrial uptake was initiated by the addition of succinate (5 mm). Release of MitoDNP-SUM was instigated by the addition of FCCP (4 μm). B) Mitochondria were incubated in the presence of antimycin A (2 μm). Two aliquots of 3 μm MitoDNP-SUM were added to the chamber, and the mitochondria were energised by hydrolysis of ATP (5 mm). Release was initiated by FCCP.