Fig. 4. HGF induces association of αDGK with Src. Quiescent cells were stimulated with HGF (250 U/ml, 15 min) and lysed in detergent-containing buffer. (A) Src was immunoprecipitated with anti-Src antibodies from either GTL 16 or PAE cell lysates. Control and anti-Src immunocomplexes were analyzed for Dgk enzymatic activity (where indicated R59949 was added in the reaction mix) (upper panel) and for Src protein by western blotting with anti-Src antibodies (lower panel). (B) Src was immunoprecipitated with anti-Src antibodies from COS cell lysates. Control and anti-Src immunocomplexes were analyzed for Dgk enzymatic activity (upper panel), for αDgk protein by western blotting with αDgk antibodies (central panel), and for Src protein by western blotting with anti-Src antibodies (lower panel). (C) Either αDgk or Src was immunoprecipitated with anti-αDgk or anti-Src antibodies from lysates of αDgk-transfected COS cells. Control, anti-αDgk and anti-Src immunocomplexes were analyzed by western blotting with anti-αDgk and anti-Src antibodies.