FIG. 6.

Mouse ZPBP1 and ZPBP2 display different release kinetics and protein processing. (A) Time course Western blot analyses of Acr-EGFP, ZPBP1, ZPBP2, and β-tubulin remaining in Acr-EGFP mouse sperm after the A23187-induced acrosome reaction (AR). β-Tubulin was used as loading control. (B and C) Western blot analysis of ZPBP1, ZPBP2, and Acr-EGFP from different fractions after the three-step (B) or the two-step (C) serial extraction of Acr-EGFP sperm. SS, sonicated supernatant; HS, high-salt extraction; RIPA, RIPA extraction; M-PER, M-PER permeabilization. (D) Western blot analysis of sperm proteins treated without (−) and with PNGase F. Anti-ZPBP1 and anti-ZPBP2 antibodies were used as shown.