Tyrosine phosphorylated Cas substrate domain affects c-crk interaction with C3G, but not endogenous FAK or Cas tyrosine phosphorylation.a) Lysates from NIH3T3 cells stably carrying vector alone or expressing the Src*/Cas(SD) chimera were immunoprecipitated with anti-phosphotyrosine (top panel), or anti-crk (middle and bottom panels) and probed with anti-HA to detect the Src*/Cas(SD) chimera (top and middle panels), or C3G (bottom panel). Lanes 1 and 2, vector and the Src*/Cas(SD) chimera transfected NIH3T3 cells respectively, whole cell extract; lanes 3 and 4, vector transfected NIH3T3 cells immunoprecipitated with anti-phosphotyrosine antibody; lane 5, NIH3T3 cells expressing the Src*/Cas(SD) chimera immunoprecipitated with anti-phosphotyrosine antibody. Arrows indicate the Src*/Cas(SD) chimera and C3G in the top, middle and bottom panels. b) A duplicate blot to that shown in (a) was probed with anti-Fak antibodies (top panel) or with anti-Cas antibodies (bottom panel). Arrows indicate FAK and Cas.