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. 2015 Sep 17;290(47):28038–28054. doi: 10.1074/jbc.M115.671115

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© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — Cellular assembly stoichiometry is not affected by the depletion of PRC2 subunit Eed. a and b, agarose gel electrophoresis analysis of nucleosomal DNAs extracted from fractions of 15 to 40% sucrose gradient. Nucleosomes were prepared from Eed^−/−/Y-Cbx4 and Eed^−/−/Y-Cbx8 (left) and Eed^+/+/Y-Cbx4 and Eed^+/+/Y-Cbx8 (right). Representative images of agarose gel are shown. Fractions 15 and 23 indicated by color-coded bars below the gels were used for single-molecule TIRF imaging. c, schematic depiction of the immobilization of YFP-PRC1·nucleosome complex on the surface by biotinylated anti-H3 antibody. d and e, percentage of fluorescence photobleaching steps of YFP-Cbx4 and YFP-Cbx8 on a nucleosome isolated from Eed^−/−/Y-Cbx4 and Eed^−/−/Y-Cbx8 (d) and from Eed^+/+/Y-Cbx4 and Eed^+/+/Y-Cbx8 (e) is shown. The color-coded bars are described in a and b. Results are means ± S.D.