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. 2011 Sep 19;194(6):855–871. doi: 10.1083/jcb.201106079

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© 2011 Moree et al.

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Figure 7. — CENP-C associates with M18BP1-1 in metaphase extract. (A) M18BP1 coprecipitates CENP-C from Xenopus egg extract. Western blot for CENP-C after immunoprecipitation (IP) of metaphase Xenopus extract with IgG or α-M18BP1. (B) CENP-C coprecipitates M18BP1 from the Xenopus egg extract. Western blot for M18BP1 after immunoprecipitation of metaphase Xenopus extract with IgG or α–CENP-C. We only detected a single M18BP1 isoform (asterisk) coprecipitating with CENP-C, which migrates at the molecular mass of isoform 1. (C) M18BP1-1 associates with CENP-C in metaphase extract. FLAG epitope–tagged M18BP1 IVT proteins were added to metaphase or interphase extracts, reactions were split in half, and M18BP1-FLAG proteins were immunoprecipitated with control (IgG) or α-FLAG (αFL) antibodies. A Western blot of the immunoprecipitates for CENP-C is shown. (D) Representative Western blots from immunoprecipitation experiments described in C. Samples were taken after incubation of M18BP1-FLAG proteins in extract. (left) α-FLAG Western blot; (right) phospho-wee1 Western blot to assess the cell cycle state of the extract. M and I indicate metaphase and interphase extract, respectively. The molecular masses of protein standards and individual proteins in kilodaltons are listed on the left of each blot.