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Fig. 1. Wee1-depletion using antisense morpholino oligonucleotides (MO). (A) MO-Wee1 was injected into both cells of a two-cell embryo. Uninjected and MO-Wee1-injected embryos were collected at various stages and lysates examined by immunoblot analysis using anti-Wee1 antibodies (* indicates a nonspecific band). (B) Two-cell embryos were injected with MO-Control, MO-Wee1 or co-injected with MO-Wee1 and WT Wee1 RNA (MO-Wee1+RNA). Embryos were collected at stage 10 and lysates examined as in A. (C) Embryos injected as in B were collected at stage 8, 10 or 12. Lysates were examined by immunoblot analysis using anti-phospho-Cdc2 or anti-Cdc2 antibodies. (D) The mitotic nuclei of injected stage 11 embryos were visualized by whole-mount immunostaining using phospho-histone H3 ({alpha}PH3). Mitotic index (n=10-12 embryos): MO-control, 8.9% (4739 nuclei); uninjected, 9.9% (1221 nuclei); MO-Wee1, 24.5% (4734 nuclei); MO-Wee1+RNA, 11.4% (2736 nuclei).