Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2006 Aug 28;174(5):665–675. doi: 10.1083/jcb.200601062

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2006, The Rockefeller University Press

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 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 4. — The Mps3 SUN domain binds to Mps2. (A) Bacterial extracts from cells expressing MBP (−), MBP-Mps2, MBP-mps2ΔC (amino acids 1–328), MBP-mps2-Ct (amino acids 328–387), and MBP-Kar1 were electrophoretically separated, and proteins were transferred to nitrocellulose membranes. Expression of MBP and the MBP fusion proteins was detected by Western blotting with anti-MBP antibodies (left). Membranes were also probed with 10 μg purified 6xHis-mps3SUN labeled with Alexa 680 in a gel overlay assay (right). (B) Nitrocellulose membranes with MBP-Mps2 were probed at 30°C with 10 μg purified 6xHis-mps3SUN (amino acids 457–617) labeled with Alexa 680 in the presence of nothing (−), 100 μg purified, unlabeled 6xHis-mps3SUN or purified, unlabeled 6xHis-mps3SUN mutants. The decrease in labeled 6xHis-mps3SUN signal was quantitated, and the signals in the presence of no competitor (−) or in the presence of the wild-type SUN domain were assigned values of 1 and 0, respectively. Mean values obtained from three independent experiments are shown. (C) Spheroplast lysates were prepared from wild-type (SLJ001), MPS2-13xMYC (SLJ1896), MPS3-3xFLAG (SLJ1898), and MPS2-13xMYC MPS3-3xFLAG (SLJ1900) strains, and the protein composition of the lysate (left) and the indicated immunoprecipitate (right) was analyzed by Western blotting with anti-MYC, anti-FLAG, anti-Cdc31, and anti-Spc29 antibodies. (D) Liquid nitrogen ground lysates were prepared from wild-type (SLJ001) and 9xMYC-MPS2 (SLJ1996) strains as described in Materials and methods. The protein composition of the lysate (left) and the anti-MYC immunoprecipitates (right) were analyzed by Western blotting with anti-MYC and anti-Mps3 antibodies.