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Figure 4. Mcm9 is required for efficient DNA replication but not Mcm2–7 loading. Egg extract prior to depletion was diluted and the indicated percentage of original extract volume analyzed by western blotting with the indicated antibodies alongside samples of control and Mcm9-depleted extracts. (B) Control-depleted and Mcm9-depleted extract were supplemented with sperm nuclei at 7.5 ng DNA/µl and α-[32P]dATP. At the indicated times, aliquots were taken, and DNA synthesis assessed by TCA precipitation and scintillation counting. The mean value with the standard error of the mean (SEM) are plotted in three sets of independently depleted mock and Mcm9-depleted extracts normalized to the maximum DNA synthesis observed in each control-depleted extract. (C) Control-depleted and Mcm9-depleted extracts were supplemented with sperm nuclei at 7.5 ng DNA/µl. Chromatin was isolated at the indicated times and immunoblotted with the indicated antibodies. A representative of three performed experiment is presented. Image published in: Gambus A and Blow JJ (2013) Copyright © 2013 Landes Bioscience. Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

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