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Xenbase Image ID: 123780

Figure 6. A nucleus is not required for pre-IC formation on chromatin. (A) Replication time-courses of single-stranded M13 DNA (pM13) or demembranated sperm nuclei (nuclei) in low speed interphase egg extracts (IEE) or high speed supernatants (HSS). (B) Chromatin was purified from control HSS (Ctl) or HSS containing 100 µM ethidium (EB) after incubation with demembranated sperm nuclei for the indicated times, or without DNA (-DNA) and western blotted against the indicated proteins. In the 20 min time-point, more material was recovered as shown by increased MCM7 signal, but the ratio of pre-IC and initiation factors to pre-RC components remained the same. (C) Assessment by immunofluorescence of the presence of pre-RC (MCM7) and initiation (PCNA and polymerase ε) complex proteins on the chromatin at 60 min in control (Ctl) HSS extracts or HSS extracts with 100 μM ethidium bromide (EB).

Image published in: Krasinska L and Fisher D (2009)

© 2009 The Author(s). Creative Commons Attribution-NonCommercial license

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