Fig. 2. αDgk kinase-inactive mutant acts as dominant negative. Quiescent PAE cells infected with either PINCOS or PINCOS/myc-αDgkK^– retrovirus were stimulated with HGF (250 U/ml, 15 min). After lysis in detergent-containing buffer, both endogenous and recombinant αDgk were immunoprecipitated with a mix of specific anti-αDgk monoclonal antibodies. The immunocomplexes were analyzed for Dgk enzymatic activity (upper panel) and for αDgk proteins, both endogenous and myc-αDgkK^–, by western blotting with anti-αDgk antibodies (lower panel).