Fig 2: The oxidative phosphorylation uncoupler FCCP reduces sperm motility and ATP content. (A–D) Motility parameters of dairy goat sperm using CASA: (A) motility tracks were generated using CASA. (B) Motility of sperm of dairy goats. (C,D) Progressive motility and straight line velocity of sperm of dairy goats. (E–G) Effect of FCCP on sperm mitochondrial functions: (E,F) opening fluorescence intensity of mPTP in mitochondria and (G) ATP content. (H–L) Expressions of COX-1, COXVB, ND3, and NRF1 determined using Western blotting after treatment with FCCP for 5 h. All results are expressed as the mean ± standard error of the mean, with asterisks indicating statistical significance for the respective control group. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. ns, no significant difference.

Fig 3: The mitochondrial translation inhibitor TIG decreases MMP and ATP levels in a dose-dependent manner. (A–C) Changes in sperm motility parameters using CASA: (A) motility of sperm of dairy goats. (B,C) Progressive motility and straight line velocity of sperm of dairy goats. (D) ATP levels in the mitochondria. (E) Mitochondrial membrane potential. (F–J) Protein levels of COX-1, COXVB, NRF1, and TFAM during 5 h of incubation with TIG determined using Western blotting. All results are expressed as the mean ± standard error of the mean, with asterisks indicating statistical significance for the respective control group. ** p < 0.01, *** p < 0.001, and **** p < 0.0001. ns, no significant difference.