FIGURE 6.
Hv1 activity affects NOX5-dependent superoxide production in human spermatozoa. A, superoxide production by spermatozoa preincubated for 5 min with or without 1 mm ZnCl2 and then stimulated with ionomycin (1 μm), PMA (1 μg/ml), or H2O2 (100 μm) was measured and expressed as described in the legend to Fig. 2. The data expressed are the mean ± S.E. of 4–6 independent experiments. Asterisks indicate statistical significance versus control cells without stimulus. **, p < 0.01; ***, p < 0.001. The number signs indicate statistical significance versus stimulated cells in the absence of ZnCl2. ##, p < 0.01; ###, p < 0.001. B, immunoprecipitation of NOX5 or HV1 protein was performed using total protein lysates isolated from sperm cells of different donors. Non-immune IgG was used to control for specificity. The content of the immunoprecipitates (IP) was analyzed by immunoblot using HV1, NOX5, and phospho-c-Abl antibodies.