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. 2013 Feb 27;8(2):e58052. doi: 10.1371/journal.pone.0058052

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© 2013 Wadelin et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Co-immunoprecipitation of endogenous histones with PRAME-FLAG-6xHis isolated from extracts of transfected HEK293 cells. Input lanes (left panels) show the presence of endogenous or FLAG-tagged proteins in extracts from cells transfected with PRAME-FLAG or empty vector. After IP with anti-FLAG antibody, immunoblots were performed with the antibodies indicated. (B) GST-pulldown experiment showing association of histones with GST or GST-PRAME proteins. Whole cell extracts of HL60 cells were incubated with immobilised GST or GST-PRAME proteins, and bound proteins separated by SDS-PAGE. Immunoblots were performed with specific antibodies to detect association of histones H2A, H2B, H3 and H4 with GST proteins. (C) Direct association of histones with GST or GST-PRAME proteins. Core histone preparations were incubated with GST beads. Following extensive washing, bound histones were separated by SDS-PAGE and revealed by western blotting using specific antibodies as indicated.