Effects of C3G and various mutant signaling molecules on adhesion of 32D cells. (A) Effects of overexpression of C3G on adhesion of 32D cells to fibronectin. 32D/EpoR-Wt cells were transfected with 30 μg of pcDNA-C3G (C3G) and 10 μg of pSG-CrkL (CrkL), as indicated, along with 1 μg of pRL-SV. The total amount of expression plasmids for each transfection was adjusted to become equal by addition of pcDNA3. Transiently transfected cells were subjected to the cell adhesion assay as described in Materials and Methods. (B) Effects of dominant negative mutants of C3G and R-Ras on adhesion of 32D cells to fibronectin. 32D/EpoR-Wt cells were transfected with 40 μg of pcDNA-C3G-dSS (C3G-dSS) or pcDNA-R-Ras43N (R-Ras43N) and 5 μg of pSG-CrkL (CrkL), as indicated, along with 1 μg of pRL-SV. Transiently transfected cells were subjected to the cell adhesion assay. (C) Effects of dominant negative mutants of Raf-1 and H-Ras on adhesion of 32D cells to fibronectin. 32D/EpoR-Wt cells were transfected with 35 μg of pcDNA-Raf-dSE (Raf-dSE) or pcDNA-H-Ras17N (H-Ras17N) and 5 μg of pSG-CrkL (CrkL), as indicated, along with 1 μg of pRL-SV. Transiently transfected cells were subjected to the cell adhesion assay.