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. 2015 Jun 15;10(8):677–691. doi: 10.1080/15592294.2015.1060387

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Figure 6. — HCF-1 and several chromatin-associated proteins but not histones bind to WGA lectin. HEK293T cells were treated with 100 µM PUGNAc or equal volume of DMSO for 24 hours as indicated and harvested to perform acid extraction of histones. The indicated soluble and chromatin-associated-proteins fractions (Left panels), and histone fraction (Right panels) were incubated for 2 hours with agarose bound WGA lectin or with agarose resin to control for non-specific binding. Two elutions (E1 and E2) of agarose and WGA bound proteins as well as the flow through (FT) fractions were analyzed by western blot using the indicated antibodies. The Coomassie Brilliant Blue staining and the anti-H2A antibody indicate that histones are found in the input and the FT fractions. Arrows indicate endogenous H2A ubiquitination (eH2Aub) and total H2A (eH2A). kDa: Molecular weight marker in kilodalton.