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. 2010 Apr 26;38(16):5409–5418. doi: 10.1093/nar/gkq262

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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).