Fig. 6

Effect of PYK2 inhibitors on fertilization. Oocytes were treated with AG17, PF04594755, AG82 (control) or DMSO (solvent control) for 45 min, then washed with Hank′s-BSA. Other groups were injected with different concentrations of GST-PERM or GST fusion proteins. The samples were fertilized, then fixed and stained with DAPI at 15 m.p.i. to assess sperm incorporation by the presence of male and female pronuclei, and at 90 m.p.i. to assess cleavage by the presence of a cleavage furrow and two somatic nuclei. An example of zygotes derived from control (DMSO) treated oocytes demonstrating the presence of two pronuclei (A, arrows) and a polar body (pb) demonstrates the criteria for sperm penetration. Panel B demonstrates the presence of a cleavage furrow and two somatic nuclei shown (B, arrows) during telophase of the second mitotic division indicating that fertilization was successful. The % of oocytes that successfully incorporated sperm (C and D, left) or cleaved to the 2-cell stage (C and D, right) are presented as the mean of > three experiments +/ S.E.M. Values represent the mean of three or more experiments +/ S.E.M. (*)=significantly different from control (P<0.05).