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. 2013 Aug 20;288(39):28293–28302. doi: 10.1074/jbc.M113.511352

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Increased dimerization and trans-autophosphorylation at Thr-283 by MEKK2. A, HA epitope-tagged MEKK2 proteins were expressed either alone or with FLAG-K385M-MEKK2 in HEK293 cells. After 24 h, HA-MEKK2 was immunoprecipitated (IP), and the co-precipitating FLAG-MEKK2 was determined by immunoblotting. B, wild-type HA-MEKK2 or HA-MEKK2 with various mutations was co-expressed with FLAG-T283A-MEKK2 or FLAG-K385M-MEKK2. HA-tagged proteins were immunoprecipitated, and the degree of Thr-283 phosphorylation and bound 14-3-3 was determined by immunoblotting and detection with the Odyssey Infrared scanner. Results are representative of three independent experiments. P, phosphorylated.