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Figure 2. Initiation times associated with different replication patterns. CHOC-400 nuclei were incubated at 10,000 nuclei/µl in egg extracts supplemented with geminin. At different times, aliquots were supplemented with 1 mM roscovitine to block further initiation events. (a) Extract was also supplemented with α-[32P]dATP at the start of the incubation. At different times thereafter, total DNA synthesis was measured by TCA precipitation and scintillation counting. (b) At different times, extract was pulse labeled with Cy3-dUTP for 5 min, and the proportion of nuclei showing different replication patterns was assessed. (c) Data from a and b were combined to provide an estimate of the times over which most initiation events associated with the different replication patterns were occurring. Image published in: Thomson AM et al. (2010) © 2010 Thomson et al. Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View |