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. 2012 Nov 5;287(53):44203–44211. doi: 10.1074/jbc.M112.365296

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Role of the ubiquitin ligase activity of Syvn on its interaction with Rer1. A, Syvn^−/− fibroblasts overexpressing Syvn-FLAG or Syvn C307A-FLAG together with hRer1 were immunoprecipitated with an anti-FLAG antibody (FLAG) or normal mouse IgG (IgG), followed by Western blot analysis of the immunoprecipitated (IP) fraction using an anti-Rer1 antibody (right panels). The expression levels of Syvn-FLAG or Syvn C307A-FLAG and Rer1 were confirmed in each lysate fraction (left panels). The antibody fraction and beads fraction served as controls and demonstrated the specificity of the immunoprecipitated bands. B, Syvn^−/− fibroblasts overexpressing Syvn-FLAG or Syvn C307A-FLAG together with hRer1 were immunoprecipitated with an anti-Rer1 antibody (Rer1) or normal rabbit IgG followed by Western blot analysis of the IP fraction using an anti-FLAG antibody (right panels). The expression levels of Syvn-FLAG or Syvn C307A-FLAG and Rer1 were confirmed in the lysate fraction (left panels). The antibody fraction and beads fraction served as controls and demonstrated the specificity of the immunoprecipitated bands.