Fig. 5. Induction of SHPS-1 tyrosyl phosphorylation by constitutive expression of MyoD in 10T
fibroblasts. (A) 10T and 10T-MyoD fibroblasts cultured in GM were subjected to immunoprecipitation with either pre-immune (PI) antisera or anti-SHP-2 specific antibodies. Immune complexes and one-tenth of the total cell lysates (TCL) were resolved and immunoblotted with pTyr antibodies. The bottom panel represents a reprobe of the above immunoblot with anti-SHP-2 antibodies. (B) Equal amounts (50 µg) of total cell lysates from 10T and 10T-MyoD cells were immunoblotted with anti-SHPS-1 antibodies. (C) 10T and 10T-MyoD cells were immunoprecipitated with either pre-immune (PI) or anti-SHP-2 antibodies and immunoblotted for SHPS-1. The bottom panel shows the immunoblot reprobed for SHP-2. (D) Lysates prepared from 10T and 10T-MyoD cells were immunoprecipitated with anti-SHPS-1 antibodies and immune complexes were immunoblotted with pTyr antibodies. (Bottom) SHPS-1 was also immunoprecipitated from a fraction of these same cell lysates and immune complexes were immunoblotted with anti-SHPS-1 antibodies as a control. The positions of molecular mass markers are shown.
