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. 2011 Dec 15;22(24):4706–4715. doi: 10.1091/mbc.E11-03-0251

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© 2011 Choo et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 6: — CSN5 binds preferentially to active CRLs. (A and B) HEK293T cells were transfected in 60-mm cell culture plates for 2 d with expression constructs for the full-length or extreme C-terminal deletion mutants of Cul2 and Cul3, as indicated at the top of each panel. The SFE mutant of Cul3 corresponds to the S53A/F54A/E55A mutant of Cul3, which is unable to bind to substrate-receptor subunits. The cells were lysed, and the lysates were subjected to V5 immunoprecipitation (IP), as described in Materials and Methods. Immunoprecipitates and aliquots of the cell lysates were analyzed by Western blotting with the indicated antibodies.