Figure 9.

Assembly of epitope-tagged mZP3, mutated at its consensus furin cleavage site, into the ZP of growing mouse oocytes. Δ-Flag-mZP3-Myc mutated at its CFCS was microinjected into growing oocytes isolated from 14-d-old mice. Oocytes were cultured for ∼50 h, and ZP were isolated in the presence of 1% NP-40 and stained with either anti-Flag or anti-Myc, followed by FITC-conjugated secondary antibody. Light (A and D) and fluorescent (B and E) images are shown for ZP treated with anti-Flag (A–C) or anti-Myc (D–F). C and F represent overlaps of A and B and D and E, respectively. Note that in B (anti-Flag), but not in E (anti-Myc), the innermost layer of the ZP is fluorescently labeled. Bar (F), 10 μm.