Xenbase Image ID: 172647

Fig. 4. RSF1 represses transcription activation from H2Aub nucleosome-containing chromatin in vitro. (A) Supercoiling assay of reconstituted chromatin template. The purified supercoiled plasmids (lane 1) were relaxed to closed circular templates using topoisomerase I (lane 2). After protease digestion and phenol-chloroform extraction, the relaxed plasmids (lane 2) were mixed with histone/ACF/Nap1 in the presence of topoisomerase I to reconstitute chromatin template that restore DNA supercoiling. DNA was then purified and changes in linking number were demonstrated (lanes 3 and 4). (B) In vitro transcription assay on naked DNA and on chromatin templates reconstituted with H2A or semisynthetic H2Aub. GAL4-VP16 and p300/AcCoA activate transcription. RSF1 inhibits transcription activation on H2Aub chromatin template, but has no effect on transcription activation on H2A chromatin. 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. Larger Image Printer Friendly View

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