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Fig. 3. (A) Expression of Myc-tagged Rho-like GTPase-mutants and HA-tagged C1199-Tiam1 in KG1. KG1 cells were retrovirally transduced with dominant negative (N19-RhoA, N17-Rac1, N17-Cdc42) or constitutively active (V14-RhoA, V12-Rac1, V12-Cdc42, L61-Rac1) GTPase mutants, or with C1199-Tiam1. Cells were lysed and expression of the mutants was detected by western blotting using anti-Myc and anti-HA antibodies, respectively. C1199-Tiam1 and the Rho-like GTPase mutants migrated according to the expected heights (RhoA > Rac1=Cdc42). (B) Rho-like GTPase mutants and C1199-Tiam1 do not alter adhesion of KG1 cells. Cells expressing either dominant negative GTPase (N19-RhoA, N17-Rac1, N17-Cdc42), constitutively active GTPase (V14-RhoA, V12-Rac1, V12-Cdc42, L61-Rac1) or C1199-Tiam1 were allowed to adhere to ALCAM-Fc-coated beads after pre-incubation with (white bars) or without (black bars) mAb AZN-L50, for 45 minutes at 37°C. Adhesion is expressed as the percentage of bound cells. Data are representative of three independent experiments.





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