Effects of mitochondrial inhibitors and the mitochondrial cofactor CoQ₁₀ on IgG-APS–induced generation of peroxides. (A) Cells were preincubated with rotenone (Rot) or antimicin A (Antim) for 1 hour, or with CoQ₁₀ for 24 hours, washed, and then stimulated with IgG-APS or IgG-NHS in the presence of the drugs. This figure shows representative flow cytometry histograms of DCF-DA fluorescence for each group of treatment. (B) Bar graph represents the mean of MFI ± SEM of 4 independent experiments. Significant differences (at P < .05) versus monocytes treated with IgG-NHS– (a) and versus IgG-APS–treated cells (b). (C) Representative fluorescent photomicrographs of ROS production by monocytes stimulated as described in panel A. Images were acquired with an LSM 5 Exciter confocal microscope (Carl Zeiss), driven by ZEN 2008 software by exciting at 488 nm to detect the DCF-DA fluorescence, and at 405 nm for 4,6-diamidino-2-phenylindole fluorescence. Samples were viewed with a EC Plan-Neofluar 20×/0.50 numerical aperture objective.