FIG. 3.

Representative in situ respiratory complex assay trace. In situ respiratory complex assays consisted of cells in a solution containing substrates for complex I and flutamide/2-hydroxyflutamide or vehicle control (shown) prior to 3 cycles of measurements and a series of compound injections into the cell culture microplate. Injections consisted of rotenone (complex I inhibitor), succinate (complex II substrate), antimycin A (complex III inhibitor), and TMPD/ascorbate (complex IV substrates) with 2 cycles of measurements following each. This series of manipulations enabled the calculation of complex I (A), II (B), and IV (C) activity. Each measurement cycle was a total of 3 min.