Figure 3.

Effect of chromanol 293B on protein tyrosine phosphorylation during human sperm capacitation. (A) Western blotting results showing the effect of chromanol 293B on protein tyrosine phosphorylation during human sperm capacitation. Sperm before capacitation incubation were used as the control group. Other sperm were treated with the vehicle control (DMSO), 20μM chromanol 293B, and 100μM chromanol 293B under capacitated conditions for 30min and 3h. Thereafter, sperm were lysed and then the proteins were resolved using 10% SDS-PAGE. Protein tyrosine phosphorylation was detected using a primary anti-phosphotyrosine antibody by Western blotting. Subsequently, the blot was stripped and probed with an anti-β-tubulin antibody as a loading control. The figure is representative of 6 separate experiments. The full uncropped immunoblots was provided in the supplementary data (Supplementary Figure 2). (B) The ratio of protein tyrosine phosphorylation levels to that of the loading control, β-tubulin. Values represent the means ± SEM of 6 experiments. For 100μM chromanol 293B group compared to vehicle control group, ^*p<0.05. (C) Indirect immunofluorescence of protein tyrosine phosphorylation in human sperm before and after capacitation for 3h. The figure is representative of 3 separate experiments.