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. 2014 Dec 30;7:39. doi: 10.1186/1756-8935-7-39

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© Stixová et al.; licensee BioMed Central. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Interaction of UBF1 with UVA-damaged chromatin. (A) Cells expressing full-length GFP-HP1β and GFP-HP1β-ΔCSD were immunoprecipitated with antibody against UBF1/2, and subsequent Western blot analysis of HP1β was performed. Quantification of immunoprecipitation fragments and original Western blots are shown. (B) FRET analysis of potential interactions between UBF1/2 (Alexa 594, red) and DNA (stained by TO-PRO-3, far-red, 642/661 nm). (Ba) Regions of interest (yellow) inspected by FRET; (Bb) quantification of UBF1/2 interaction with DNA in the nucleolus and nucleoplasm of irradiated and non-irradiated cells. FRET efficiency is shown in percentage ± standard error. For each event, 10 nuclei were analyzed in three independent experiments. (C) Immunoprecipitation verified the interaction between HP1β and UBF1/2; HP1β and H3K9me3 were used as reference interacting partners. Immunoprecipitation was performed in the presence or absence of EtBr; 20 μg of protein lysate was used as input. IF, immunofluorescence; irrad., irradiated; Nu, nucleolus; Nupl, nucleoplasm.