Extended Data Fig. 7: Roles of MMP and ROS in mitopherogenesis.
(a–c) Microscopic images and corresponding bar graph showing that N2 spermatids treated with FCCP can still be activated to form mature spermatozoa in the presence of pronase (a,b). The activation rate of the sperm from the solvent control and the FCCP treatment group are 100% and 99%, respectively (c). This result supports the notion that the inhibition of mitopherogenesis by FCCP shown in Fig. 5c is not a result of reduced spermatid viability. Each dot represents the percentage of mature spermatozoa in one experiment. 200 cells from 3 biologically independent experiments were analysed. Green arrow, spermatozoa. (d) Box-and-whisker graph indicating that antioxidants treatment showed no effect on basal level of mitopherogenesis and pronase-induced mitopherogenesis. The antioxidants, tempol that scavenges superoxide anion O2− and acetylcysteine (NAC), mainly scavenging hydrogen peroxide H2O2 and hydroxyl radical OH−, were used to reduce ROS levels in the isolated spermatids, followed by treatment with/without pronase. The number of mitophers were analysed by FACS. n = 25,000 spermatids per group from 5 biologically independent experiments were analysed. Source numerical data (c,d) are available in source data.
