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. 2014 Apr 30;4(4):140043. doi: 10.1098/rsob.140043

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© 2014 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 1. — Eic1 and Eic2 are Mis18-interacting proteins. (a) LC-MS/MS analysis of Myc-tagged Mis16^{RbAp46/48/Hat2} and Mis18 immunoprecipitates from S. pombe whole cell extracts identifies two previously uncharacterized proteins, Eic1 (SPBC27B12.02) and Eic2 (SPBC776.16). Average number of unique peptides reproducibly identified from three independent experiments is shown. (b) Primary sequence alignments of Eic1 (top) and Eic2 (bottom) orthologues identified among Schizosaccharomyces species. (c) Eic1-GFP co-immunoprecipitates with both Mis16-Myc and Mis18-Myc, while Eic2-GFP only co-immunoprecipitates with Mis18-Myc. The asterisk (*) in the bottom panel denotes the IgG heavy chain. (d) Eic1 directly interacts with Mis18 and Mis16. 6xHis-Eic1 was co-expressed with GST alone, GST-Mis18 or GST-Mis16 in E. coli. Coomassie-stained SDS-PAGE gels showing reciprocal GST (lanes 1–3) and His (lanes 6–8) pulldowns from E. coli lysates are shown. Also shown are reciprocal pulldowns of 6xHis-Mis16 co-expressed with GST alone or GST-Mis18 (lanes 4–5 and 9–10).