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. 2013 Feb 1;41(5):3162–3172. doi: 10.1093/nar/gkt043

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© The Author(s) 2013. 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/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Analysis of the role of ORC4R in pre-RC formation. (A) ATPase activity of ORC and ORC4R in the absence (columns 1 and 2) and presence of DNA (columns 3 and 4). (B) The stability of pre-RCs formed in the presence of ORC (lanes 1–6) or ORC4R (lanes 7–12). Pre-RCs were formed in the presence of low salt buffer (lanes 1 and 7) or with buffer containing 100, 200, 300, 400, 500 mM sodium chloride, respectively (lanes 2–6 and 8–12). (C) Pre-RC complexes were crosslinked in solution. Pre-RC ATP (lanes 1, 5, 6, 10, 11, 15, 16 and 20), pre-RC ATPγS (lanes 2, 7, 12 and 17) and pre-RC with ORC4R (lanes 3, 4, 8, 9, 13, 14, 18 and 19). (D) Electron micrographs of metal-shadowed pre-RCs formed with ORC (upper picture) and ORC4R (lower picture). (E) Electron micrographs of uranyl acetate-stained pre-RC samples prepared with ORC (upper picture) and ORC4R (lower picture).