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. 2011 Feb 22;286(16):13954–13965. doi: 10.1074/jbc.M111.220103

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — A glycine-zipper motif in the TM domain of EWI-2/EWI-2wint is essential for the interaction with CD81. A, schematic representation of the different EWI-2 mutants. Expression of EWI-2 (wt) in CHO cells leads to the production of EWI-2 and EWI-2wint. Residues in the glycine-zipper or glycine-rich motif of EWI-2 were replaced by leucines. FLAG-tagged proteins were co-expressed with CD81 in CHO cells, biotinylated, lysed in PBS/Brij97/EDTA, and immunoprecipitated with 5A6 or M2. Proteins were resolved by SDS-PAGE under nonreducing conditions and detected using HRP-streptavidin.