Slit-Robo signaling recruits Hakai to E-cad and induces E-cad ubiquitination. (A-D) Hakai binding to E-cad in HCT116/V, HCT116/Robo1, HCT116/Robo1/Slit2, SW620/V and SW620/shRobo1 cells in the absence or presence of mIgG, R5, PBS and hSlit2. E-cad was immunoprecipitated from the cell lysates followed by immunoblotting with the Abs to E-cad and Hakai. (E-H) E-cad ubiquitination in HCT116/V, HCT116/Robo1, HCT116/Robo1/Slit2, SW620/V and SW620/shRobo1 cells in the absence or presence of mIgG, R5, PBS and hSlit2. E-cad was immunoprecipitated from the cell lysates followed by immunoblotting with the ubiquitin and E-cad Abs. Ub, ubiquitin. Results are representative of at least three independent experiments.