Autokinase assay and phosphorylation on tyrosine of the CEP110-FGFR1 fusion protein.

NIH3T3 cells transiently transfected by either CEP110-FGFR1 cDNA (CEP110-FGFR1) or the empty vector (pcDNA3) and FGFR1 overexpressing cells (NFlg26) were cultured in the presence (+) or absence (−) of FGF1 plus heparin. Immunoprecipitates using anti-C-FGFR1 antibody were analyzed for autokinase activity (left panel) (autoradiography of 4 hours for all samples) and phosphorylation on tyrosine after Western blot with anti-phosphotyrosine antibody 4G10 (right panel). The position of molecular mass standards (in kd) is indicated.