WWP2 binds to N3-ICD but not to N1-ICD, N2-ICD, or N4-ICD.

<p>(<b>A</b>) Immunoprecipitation (IP) using either anti-FLAG or anti-V5 antibody was carried out using extracts prepared from 293T cells transfected with ICD construct alone or in combination with WWP2 as indicated. The interaction of FLAG-tagged WWP2 with the different V5-tagged ICD was evaluated by western blot (WB) with the respective antibody. Only N3-ICD was co-immunoprecipitated with WWP2. (<b>B</b>) Schematic representation of N1-ICD and N3-ICD, along with Y-A mutant of N3-ICD and N1/N3-ICD chimera. (<b>C</b>) Immunoprecipitation using either anti-FLAG or anti-V5 beads was carried out using extracts prepared from 293T cells transfected with constructs alone or in combination as indicated. The interaction of FLAG-tagged WWP2 with the different V5-tagged ICD was evaluated by western blot (WB) with their respective antibodies. (<b>D</b>) Schematic representation of WWP2 and a construct coding for the four WW domains (WWD). (<b>E</b>) Immunoprecipitation using either anti-FLAG or anti-V5 beads was carried out using extracts prepared from 293T cells that were transfected with constructs alone or in combination as indicated. The interaction of FLAG-tagged WWD with the different V5-tagged N-ICDs was evaluated by western blot (WB) with their respective antibodies. Result shows that interaction between N3-ICD and WWP2 is mediated by the WW domain.