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. 2018 Aug 6;9:3103. doi: 10.1038/s41467-018-05581-y

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — HIRA homooligomerizes in cells and forms a homotrimer in vitro. a YFP constructs of human HIRA and mutants. The WD40 repeat (aa 1–369), B (aa 439–475) and C (aa 763–963) domains involved in UBN, ASF1a, and CABIN1 interactions, respectively, are shown. Star indicates single amino acid substitution with alanine. The co-immunoprecipitation efficiency of endogenous HIRA or HIRA-HA (HIRA coIP) is indicated for each construct, “+” indicates that the efficiency of coIP is similar to the one obtained with the wt HIRA construct and “−” indicates that the efficiency of the coIP is decreased. b Western blot analysis of anti-GFP-immunoprecipitates from U2OS nuclear extracts expressing YFP-tagged proteins. c Western blot analysis of anti-GFP-immunoprecipitates from U2OS nuclear extracts expressing both YFP-tagged and HIRA-HA proteins. d Western blot analysis of anti-GFP-immunoprecipitates from U2OS nuclear extracts expressing YFP-tagged proteins. In b, c and d, input corresponds to 10% of nuclear extract used for each experiment. e Equilibrium sedimentation of recombinant proteins HIRA(661–1017), CABIN1 full length (FL) and HIRA(661–1017) + CABIN1 FL. Theoretical (open symbols) and experimental (closed symbol) curves are shown