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Fig. 4. The intra-S-phase checkpoint reduces the number of active replication forks. (A,B) Sperm nuclei were incubated at 10 ng DNA µl^-1 in interphase extract with or without 7.5 µM or 15 µM aphidicolin, minus (A) or plus (B) 5 mM caffeine. During early S phase (35 minutes), nuclei were isolated and transferred to fresh extract supplemented with [-³²P]dATP and 0.5 mM roscovitine. Total DNA synthesis was measured at the indicated times after transfer (1-30 minutes). (C,D) Sperm nuclei were incubated in interphase extract at 10 ng DNA µl^-1 supplemented with 40 µM aphidicolin minus (C) or plus (D) caffeine. Sperm chromatin was incubated for 35 minutes, 45 minutes or 60 minutes and was then isolated and transferred to fresh extract supplemented with [-³²P]dATP and 0.5 mM roscovitine. Total DNA synthesis was measured at different times after transfer.

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