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. 2009 May 13;37(11):e81. doi: 10.1093/nar/gkp273

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© 2009 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — Supercoiling assay: purified hyper- and hypo-phosphorylated H3/H4 and core histones are functional and form tetrasomes and octasomes in the presence of chromatin assembly factors. Topo I-relaxed BlueScript plasmid was incubated with the respective histone fractions, which were pre-incubated with Q-Sepharose-isolated nucleosome assembly fraction (QS500) from HeLa cells. The topoisomer distribution in the plasmid population was analyzed by 0.9% agarose gel electrophoresis following proteinase K treatment and phenol/chloroform/isoamyl alcohol extraction. Reconstitution with M-phase hyper-phosphorylated H3/H4 histones (A) or core histones (B). Histones isolated from logarithmically proliferating cells, hence hypophosphorylated, were used in parallel. The components of the reaction mixtures and the histones used are indicated at the top of the figure.