FIGURE 5.

ZP or PNU282987 increase EGFR phosphorylation, which is Src-mediated. A, Sperm from WT (Cont) or α7-null mice (MUT) were incubated in capacitation medium for 1.5 h. B, PP1 (5 μm) or α-BgT (100 nm) was added for the last 30 min of incubation. A, EGF (1 ng/ml), ZP (∼7.5 ZP/μl), or PNU (in A and B, 5 μm) was added for an additional 5 min. Sperm proteins were extracted with SDS lysis buffer as described under “Experimental Procedures.” The extracts were separated on SDS-PAGE and transferred to nitrocellulose membrane that was incubated with anti-phospho-EGFR (Tyr-845) and with anti-tubulin for protein amount. The data represent one experiment typical of three repetitions performed.