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FIGURE 6. NSD2 complex dimethylates H3K36 on nucleosomes or octamers plus DNA. A, NSD2 complex was purified from HT1080 cells stably expressing FLAG- and Myc-tagged NSD2 by anti-FLAG resin. As a control, a similar purification from a stable cell line with empty vector was carried out, side by side (Mock). The input (IN), flow though (FL), and eluate (E) from the FLAG purification were separated by SDS-PAGE and analyzed by Western blots (left panel) using anti-FLAG M2 or anti-NSD2 antibodies as indicated at the bottom and by silver staining (right panel). B and C, HKMT assay performed with the isolated NSD2 complex and nucleosomes composed of wild type (WT), H3K36A, or premethylated H3K36 as indicated (B, top panel) or with octamers as a function of increasing amounts of 41-bp dsDNA (C, top panel). The asterisk denotes a proteolyzed form of H3. CBB staining of histones is shown (bottom panels). Image published in: Li Y et al. (2009) © 2009 by The American Society for Biochemistry and Molecular Biology, Inc. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license Larger Image Printer Friendly View

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