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. 2016 Jan 8;291(10):5428–5438. doi: 10.1074/jbc.M115.703637

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Rco1 forms a homodimer within Rpd3S. A, co-immunoprecipitation showing that differently tagged Rco1 proteins associate with each other in yeast. Whole cell extracts (WCE) were prepared from yeast strains YBL766 (Rco1-TAP), YBL770 (Rco1-HA), and YBL772 (Rco1-TAP/Rco1-HA) and then immunoprecipitated (IP) using IgG-Sepharose before Western blotting. B, recombinant Rpd3S complex contains two copies of Rco1 that are independent of CHD and PHD1. FLAG and HA tandem purification was carried out for indicated Rpd3S complexes. CBB, Coomassie Brilliant Blue; WB, Western blot. C and D, Rco1 forms a homodimer in the absence of Eaf3 in a baculovirus expression system. E, an updated model of the Rpd3S-interaction network.