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. 2012 Jun 8;287(32):26840–26853. doi: 10.1074/jbc.M112.355743

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

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FIGURE 13. — H147N/H149N/H151N-imAPP is overubiquitinated compared with WT-imAPP. Cell lysates of HEK293 cells stably expressing WT- or H147N/H149N/H151N-APP or the vector alone were immunoprecipitated (IP) and immunoblotted (WB) with WO2 antibody. A, FK2 antibody (which recognizes polyubiquitin and monoubiquitin conjugates) immunoprecipitation showed that only imAPP bearing the H147N/H149N/H151N mutation (not WT-imAPP) is ubiquitinated. B, positive control. WO2 antibody immunoprecipitation showed the usual doublet for WT-APP and only one band for H147N/H149N/H151N-APP. C, negative control. Mock immunoprecipitation only showed nonspecific bands. Note that blots in A and C are a result of longer signal time collection than the blot in B.