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

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© 2011 Moree et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Characterization of Xenopus M18BP1. (A) Western blot of Xenopus egg extract with preimmune sera (Preimmune) or affinity-purified rabbit antibody raised against Xenopus M18BP1 (α-M18BP1). α-M18BP1 recognizes both isoforms of xM18BP1, and both isoforms of xM18BP1 are present in egg extract. The asterisk denotes a cross reacting band. (B) M18BP1 is not present on Xenopus sperm chromatin. After decondensation with Xenopus Nap1, Xenopus sperm chromatin shows staining for CENP-A but not M18BP1. (C) Time course of M18BP1 localization in the Xenopus egg extract. Xenopus sperm chromatin was incubated in metaphase extract for 30 min, and sperm chromatin was stained for M18BP1 at various time points after release from metaphase arrest. Time (minutes) after release from metaphase is indicated above each image column. M18BP1, CENP-A, or merged M18BP1, CENP-A, and DNA staining are indicated on the left. (D) M18BP1-1 assembles at sperm centromeres in metaphase extract, and both M18BP1 isoforms assemble at sperm centromeres in interphase extract. Metaphase extracts are shown on the left, and interphase extracts are shown on the right. The FLAG epitope–tagged M18BP1 isoform that was added to the extract is indicated on the left. Immunostaining is shown above the images. Bars, 10 µm.