Figure 3
Overexpression of Wee1 delays the MBT and alters Cdk2. Embryos were injected at the one-cell stage with 2.5 ng of Wee1 or luciferase (control) mRNA. (A, B) Embryos were collected at the indicated stages and lysates were subjected to Western analysis using (A) Cdc25A and (B) cyclin E antibodies. Approximate timing of the MBT in luciferase expressing control embryos is indicated. (C) Embryos were collected at 4 hrs pf and lysates were immunoprecipitated with cyclin E serum. Immunoprecipitates were subjected to Western analysis using a primary phosphoCdk antibody (Cell Signaling Technology). (D) Cyclin E immunoprecipitates were collected at the times indicated and assayed for Cdk2 activity by phosphoryation of histone H1. The amount of γ-32P ATP incorporated in histone H1 is expressed as radioactive counts per minute (CPM).