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Fig. 3. Aphidicolin induces a caffeine-sensitive replication checkpoint. (A-D) Sperm nuclei were incubated at 15 ng DNA µl^-1 in interphase egg extract supplemented with [ ${alpha}$ -³²P]dATP plus various concentrations of aphidicolin plus or minus 5 mM caffeine. (A) Total DNA synthesis was measured at 120 minutes. (B) DNA synthesis was measured between 15 minutes and 120 minutes as indicated. (C,D) At 120 minutes, DNA was separated on an alkaline agarose gel and autoradiographed. The migration of end-labelled ${lambda}$ -phage HindIII-digested DNA is also shown. (E,F) Sperm nuclei were incubated at 15 ng DNA µl^-1 for 35 minutes in interphase egg extract supplemented with 7.5 µM aphidicolin. The reaction was split in two and supplemented with 0.5 mM roscovitine minus (E) or plus (F) 5 mM caffeine. At 5 minute intervals, aliquots were pulse labelled for 2 minutes with [ ${alpha}$ -³²P]dATP and then DNA was separated on an alkaline agarose gel and autoradiographed. The migration of end-labelled ${lambda}$ -phage HindIII-digested DNA is also shown.

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