Fig. 6. Inducible activation of MyoD in 10T
MyoD-ER fibroblasts promotes SHPS-1 tyrosyl phosphorylation and association with SHP-2. 10T MyoD-ER cells were cultured in GM or DM in the absence (-E) or presence (+E) of estradiol (0.1 µM) for 18 hours. Cell lysates prepared from these cultures were (A) immunoblotted with anti-MHC antibodies or (B) immunoprecipitated with anti-SHPS-1 antibodies followed by pTyr immunoblotting (top). As a control an equal fraction of these lysates were immunoprecipitated for SHPS-1 and immunoblotted with anti-SHPS-1 antibodies. Gab-1 was also immunoprecipitated under the indicated conditions and subjected to anti-pTyr immunoblotting (bottom). (C) SHP-2 was immunoprecipitated under the conditions indicated; tyrosyl phosphorylation of the p120/SHPS-1 complexed with SHP-2 was assessed by immunoblotting with anti-pTyr antibodies (top) and the amount of SHPS-1 in the complex was determined by immunoblotting for SHPS-1 (middle). The bottom panel shows the SHP-2 reprobe as a control.
