Figure 5.
TrkA coprecipitates with ephrinA5FLAG in PC12 cells after NGF stimulation. PC12 cells transiently transfected with ephrinA5FLAG or with phosphorylated enhanced yellow fluorescent protein (eYFP) (as a control of transfection efficiency) were treated with 50 ng/ml NGF for several time intervals. A, After immunoprecipitation of ephrinA5FLAG, the presence of TrkA in the precipitates was analyzed by Western blot, and the precipitation of ephrinA5 was verified. IB, Immunoblot; IP, immunoprecipitation. B, Total cell lysates from the same samples were run to check NGF-induced activation of signaling cascades with anti-phospho-Akt and anti-phospho-Erk1/2 antibodies, and, after membrane stripping, protein loading was checked with anti-tubulin (bottom panels) antibodies.