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Figure 3. Cdc6 halts DNA replication before Cdk2-dependent steps. (A) A schematic representation of the experimental procedures. (B) Sperm nuclei (6 ng DNA/µl) were incubated with extracts containing aphidicolin (40 ng/µl) for 60 min and isolated to obtain aphidicolin nuclei. Amount of DNA synthesized was measured during a 90-min incubation after untreated sperm chromatin (sp) or aphidicolin nuclei (ap) were added in extracts containing buffer (none), aphidicolin (aph; 40 ng/µl) or GST–Cdc6 (700 nM). DNA synthesis is represented as a percentage of the radioactivity incorporated into DNA in the samples of interest to that after incubation with control extracts for 90 min. (C) The same experiment as shown in B but using p21 (5 ng/µl) instead of aphidicolin for nuclear preparation to obtain p21 nuclei (21) and for additives of the second incubation (p21). Image published in: Kundu LR et al. (2010) © The Author(s) 2010. Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

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