Figure 5.
CDK1 protein is newly synthesized after fertilization. [35S]-methionine labeled proteins from embryos collected at the indicated times after fertilization were incubated on p13suc1-sepharose beads; affinity-purified proteins were resolved on SDS-PAGE and transferred to a nitrocellulose membrane. After exposure on a PhosphorImager screen (autoradiography), the membrane was incubated with anti-PSTAIR antibodies directed against the CDK1 protein (WB). Autoradiography and western blot on total proteins are shown in the bottom panel. Fertilized (left) and PP242-treated (right) embryos are shown. As a control, [35S]-methionine labeled embryos were cultured in presence of emetine, which was added 5 min after fertilization to inhibit protein synthesis, and the lysate prepared from embryos harvested at 90 min was used for p13suc1-sepharose bead purification (F+emet). The experiment was performed twice.