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. 2009 Nov 23;19(4):573–583. doi: 10.1093/hmg/ddp524

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© The Author 2009. 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/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Huntingtin associated with PRC2 in the nucleus. (A) Fluorescent microscope images of an immunostained section of a day 4 Hdh⁺/Hdh⁺ wild-type (WT) EB, demonstrating the nuclear conformation of huntingtin (Htt), detected by AP194 antibody (8), and anti-Ezh2-signal in DAPI-stained nuclei (merge). (B) Immunoblot showing co-immunoprecipitation of Ezh2 and Suz12 with huntingtin (Htt), from nuclear extracts of day 4 Hdh⁺/Hdh⁺ (WT) EBs. (C) Plot of the results of chromatin immunoprecipitation analysis of Hoxb9, demonstrating enrichment of huntingtin (Htt) and trimethylated histone H3K27 (H3K27me3) in day 4 Hdh⁺/Hdh⁺ wild-type (WT) EBs, that is not apparent in the absence of huntingtin in Hdh^ex^4/5/Hdh^ex^4/5 (KO) EBs, but is increased in Hdh⁺/Hdh^Q¹¹¹ knock-in (KI) EBs, expressing 111-glutamine huntingtin (n = 3; anti-Htt KO versus WT P < 0.0004; anti-histone H3K27me3 KO versus WT P < 0.0037; anti-Htt KI versus WT P < 0.4267; anti-histone H3K27me3 KI versus WT P < 0.0157).