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. 2011 Jul 20;31(29):10602–10614. doi: 10.1523/JNEUROSCI.0436-11.2011

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Copyright © 2011 the authors 0270-6474/11/3110602-13$15.00/0

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Figure 4. — The JM1 domain of TrkB interacts directly with JIP3. A, Schematic representation of the TrkB mutants generated. HEK293 cells were transiently cotransfected with Flag-tagged TrkB-FL or TrkB mutants and HA-tagged JIP3. Immunoprecipitation was performed using rabbit anti-HA antibodies, and immunoblotting was performed with mouse anti-HA or mouse anti-Flag antibodies. B, Schematic representation of the TrkB juxtamembrane domain, IL2R, and chimeric IL2R–BJM mutants. HEK293 cells were transiently transfected with the indicated constructs. Immunoprecipitation was performed using mouse anti-Flag antibodies, and immunoblotting was performed with rabbit anti-HA or rabbit anti-Flag antibodies. C, Coimmunoprecipitation of JIP3 with TrkB-FL, TrkA-FL, and TrkC-FL. HEK293 cells were cotransfected with Flag-tagged Trks and HAJIP3; immunoprecipitation was performed using mouse anti-Flag antibodies, followed by immunoblotting with rabbit anti-Flag or rabbit anti-HA antibodies. Multiple sequence alignment on the JM1 domain of three Trk receptors was performed by ClustalW2. D, TrkB-JM1 confers TrkA the binding ability with JIP3. A schematic representation of TrkA and chimeric TrkA-BJM1 mutant is shown. Immunoprecipitation was performed using mouse anti-Flag antibodies, and immunoblotting was performed with rabbit anti-HA or rabbit anti-Flag antibodies.