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. 2014 Aug 20;289(41):28448–28459. doi: 10.1074/jbc.M114.561241

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Specificity of the interaction between Fbxo45 and N-cadherin. a, 293T cell line overexpressing streptavidin/HA-tagged Fbxo45 (F45) was generated and cell lysate (CL) from the cell line were subjected to IP with streptavidin resin followed by Western blotting with indicated antibodies. b, HT-1376 cells were transfected with streptavidin/HA-tagged Fbxo45 (F45) with or without N-cadherin as indicated and CL were subjected to IP with streptavidin resin followed by Western blotting with indicated antibodies. c, the CL from the Fbxo45-overexpressing cell line were subjected to IP with anti-MYCBP2 antibody followed by Western blotting with indicated antibodies. d, exclusive interaction of N-cadherin (CDH2) with Fbxo45. 293T cells were transfected with empty vector (EV) or the indicated streptavidin/HA-tagged (Fbxo45, Fbxw1, Fbxl1) and FLAG-tagged (Fbxw2, Fbxw4, Fbxw5, Fbxw7, and Fbxo22) F-box protein constructs. CL were subjected to IP with anti-FLAG or streptavidin/HA resin followed by Western blotting with indicated antibodies. e, reciprocal interaction of N-cadherin with Fbxo45. Whole cell lysate (WCL) from 293T cells stably expressing streptavidin/HA-tagged Fbxo45 were subjected to IP with anti-CDH2 antibody followed by Western blotting with the indicated antibody.