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. 2011 Jul 8;39(19):8329–8341. doi: 10.1093/nar/gkr529

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© The Author(s) 2011. 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 4. — Enrichment of active modifications in H3.3 nucleosomes. (A) Mononucleosomes containing ectopic H3 or H3.3 were prepared as in Figures 1A and S1A, and analyzed by western blotting using antibodies that recognize H3–K4 methylation (α-H3K4me1/me2/me3) H3-K9 methylation (α-H3K9me1/me2/me3), H3–K27 methylation (α-H3K27me1/me2/me3), H2A acetylation (α-H2Aac), H2B acetylation (α-H2Bac), H3 acetylation (α-H3ac) and H4 acetylation (α-H4ac). Lanes 1, 4, 7 and 10, mock-purified materials; lanes 2, 5, 8 and 11, H3 mononucleosomes; lanes 3, 6, 9 and 12, H3.3 mononucleosomes. (B) Cells were mock-treated (−) or heat-treated (+) for 30 min, and modification status of promoter nucleosomes was determined. ChIP assays were essentially as described in Figure 3D.