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Fig. 2. Inhibition of CDK activity prevents later origins from firing. (A) Sperm nuclei were incubated at 15 ng DNA µl^-1 in egg extract supplemented with [-³²P]dATP. Aliquots were supplemented with 0.5 mM roscovitine at 20, 25, 30, 35, 40 and 50 minutes. Samples with no added roscovitine are shown by open squares. At the indicated times, samples were assayed for total DNA synthesis. (B) Sperm nuclei were incubated at 15 ng DNA µl^-1 in egg extract supplemented with biotin-dUTP. 0.5 mM roscovitine was added at the indicated times. At 120 minutes, nuclei were isolated and stained with Texas Red streptavidin to reveal nuclei that had undergone DNA replication. The proportion of biotin-positive nuclei for each point is shown. (C,D) Sperm nuclei were incubated at 15 ng DNA µl^-1 in egg extract. At 35 minutes (C) or 45 minutes (D), aliquots were supplemented with 0.5 mM roscovitine. Samples were pulse labelled with [-³²P]dATP for 2 minutes at the indicated times. DNA was separated on an alkaline agarose gel and autoradiographed. The migration of end-labelled -phage HindIII-digested DNA is also shown.

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