Figure 6.

Inhibition of the PRDX6 system increases ROS levels. Spermatozoa were incubated for 2 h with different concentrations of the inhibitors of (A) the 2-Cys PRDXs, (B) the TRX/TRD system, (C) the NADPH-suppliers enzymes, (D) the reactivation system of the PRDX6 peroxidase activity or (E) the Ca²⁺-iPLA₂ activity of PRDX6, then washed and added 10 μM H₂DCFDA for 30 min at 37°C. Control positive (C+) was done by incubating spermatozoa with 100 μM of ferrous perchlorate (II) and 1 mM of H₂O₂ under the same conditions. The mean fluorescence of label cells was analyzed by MACSQuant Analyzer flow cytometry and results are presented relative to control without inhibitors (C−) as the mean ± SEM of six measurements performed with sperm samples from different donors. *A value significantly (P ≤ 0.05) different from the control (C−) (ANOVA and Bonferroni post hoc test).