Figure 2.

Roles of ezrin and RhoGDI1 in actin polymerization during capacitation. (A): Western blot analysis of human sperm lysates extracted before and after capacitation for 2.5 or 5 h. The blots were probed with rabbit anti-phospho-ezrin (Thr₅₆₇)/radixin (Thr₅₆₄)/moesin (Thr₅₅₈) antibody (p-erzin). (B): Differential expression of RhoGDI1 before and after capacitation, as analyzed by two-dimensional electrophoresis analysis. The gels were stained with the silver or Pro-Q Diamond phosphoprotein gel stain kit. The graph (bottom) depicts the quantification of the silver-stained spots. ^*P < 0.01, compared with before capacitation. (C): Co-immunoprecipitation assay with ezrin and RhoA antibodies for pull down and RhoGDI1 antibody for detection. (D): Comparison of F-actin expression in human sperm before and after capacitation using Texas Red-conjugated phalloidin staining. Cap, capacitation; DIC, dynein intermediate chain; Pre, before capacitation. Bar = 10 μm.