Figure 2.

PRDXs are involved in the regulation of tyrosine phosphorylation induced by FCSu during human sperm capacitation. (A) Percoll-selected spermatozoa were capacitated with 10% FCSu in BWW medium at 37°C for 4 h in the absence (None) or presence of 5 or 10 μM TSP or MJ33 added at different times of incubation. (B) Effect of 10 μM TSP and MJ33 on spermatozoa incubated with or without FCSu for 4 h at 37°C. Sperm suspensions were supplemented with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer and solubilized proteins from 0.1 × 10⁶ spermatozoa were loaded in each well, electrophoresed, electrotransferred and immunoblotted using the anti-P-Tyrosine antibody (P-Tyrosine; 1:10 000). Equal loading in each well was determined by silver staining the membranes or re-blotting with anti-α-tubulin antibody (1:10 000). The immunoblots presented are representative of four experiments (n = 4 different donors). One sperm sample was used for all the experimental lanes shown on the immunoblots. Relative intensity of phosphor tyrosine in A was normalized to the relative intensity of α-tubulin of each sample and expressed as mean ± SEM. *P < 0.05 versus all other groups using ANOVA and Bonferroni test (n = 4).