Figure 5
Figure 5. Kpm/Lats2 can associate with YAP2 via its first WW domain and phosphorylate YAP2, but it requires not this association but kinase function of Kpm/Lats2 to stabilize p73. (A) Any one of HA-tagged Kpm-wild type (wt), Kpm-kinase dead (kd), or mock was cotransfected with any one of FLAG-tagged YAP2-wild type (wt), YAP2-S127A (mutant form S127 to A; S127 is Akt-phosphorylation site), YAP2-1WW* (mutant form of first WW domain), YAP2-2WW* (mutant form of second WW domain), or mock into 293T cells by the calcium phosphate method. Upper lanes represent Western blotting with anti-HA in cell lysates. Middle lanes and lower lanes represent Western blotting with anti-FLAG and anti-HA, respectively, in the immunoprecipitates by anti-FLAG. Three independent experiments were performed and representative data are shown. (B) The coimmunoprecipitate fraction by anti-FLAG from each lysate was treated with or without CIP and then analyzed by Western blotting with anti-YAP polyclonal antibody and anti–phospho-YAP (Ser127) polyclonal antibody. Arrow indicates mobility shift band of YAP2. Three independent experiments were performed and representative data are shown.

Kpm/Lats2 can associate with YAP2 via its first WW domain and phosphorylate YAP2, but it requires not this association but kinase function of Kpm/Lats2 to stabilize p73. (A) Any one of HA-tagged Kpm-wild type (wt), Kpm-kinase dead (kd), or mock was cotransfected with any one of FLAG-tagged YAP2-wild type (wt), YAP2-S127A (mutant form S127 to A; S127 is Akt-phosphorylation site), YAP2-1WW* (mutant form of first WW domain), YAP2-2WW* (mutant form of second WW domain), or mock into 293T cells by the calcium phosphate method. Upper lanes represent Western blotting with anti-HA in cell lysates. Middle lanes and lower lanes represent Western blotting with anti-FLAG and anti-HA, respectively, in the immunoprecipitates by anti-FLAG. Three independent experiments were performed and representative data are shown. (B) The coimmunoprecipitate fraction by anti-FLAG from each lysate was treated with or without CIP and then analyzed by Western blotting with anti-YAP polyclonal antibody and anti–phospho-YAP (Ser127) polyclonal antibody. Arrow indicates mobility shift band of YAP2. Three independent experiments were performed and representative data are shown.

Close Modal

or Create an Account

Close Modal
Close Modal