Fig. 4. SHP-2 associates with Gab-1 in C2C12 myoblasts. (A) Lysates prepared from C2C12 myoblasts cultured in GM were immunoprecipitated with either preimmune (PI) antisera or anti-SHP-2 antibodies. Immune complexes were immunoblotted with anti-Gab-1 antibodies. The bottom panel is a reprobe with anti-SHP-2 antibodies from the above experiment. (B,C,D) Tyrosyl dephosphorylation of Gab-1 and constitutive association with SHP-2 during C2C12 myogenesis. Lysates prepared from C2C12 myoblasts were cultured in either GM or DM for the indicated times. (B) Total cell lysates were immunoblotted with anti-Gab-1 antibodies. Lysates prepared from C2C12 myoblasts under the indicated conditions were also subjected to immunoprecipitation with either (C) anti-Gab-1 antibodies followed by pTyr immunoblotting or (D) immunoprecipitation with anti-SHP-2 antibodies followed by anti-Gab-1 immunoblotting. The bottom panel in C represents an equal fraction of the lysates from the experiment above that was immunoprecipitated for Gab-1 and immunoblotted with anti-Gab-1 antibodies as a control. Bottom panel in D is a reprobe with anti-SHP-2 antibodies of the above experiment. The positions of molecular mass markers are shown.
