Fig. 1. RSF1 preferentially associates with H2Aub-enriched nucleosomes in HeLa cells expressing HA-ubiquitin. (A) Silver staining of a protein gel containing equal amounts of H2Aub-enriched or -depleted mononucleosomes reveals polypeptides enriched specifically within each group of nucleosomes (arrowheads). Different staining times were used for core histones and the rest of the proteins. (B) Anti-HA immunoblot of a protein gel containing an aliquot of the above samples shows that a single protein corresponding to the H2Aub position containing HA-ubiquitin (arrowhead). (C) RSF1 is enriched in the H2Aub-enriched nucleosomes in the SILAC assays, shown by log2-fold enrichment of RSF1 abundance, quantified by the intensity of a RSF1 unique peptide (Fig. S1), in H2Aub-enriched (+ub) or -depleted (−ub) nucleosomes. H and L represent heavy and light lysine labeling, respectively. (D) Immunoblot analyses of proteins contained in the flow-through (Ft) or the eluate (E) of anti-HA IP of H2Aub-containing nucleosomes show that RSF1 is enriched in the H2Aub nucleosomes, along with SNF2H, another component of the RSF complex. In, input. Antibodies used are indicated at the left.
Image published in: Jones AE et al. (2017)
Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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