Fig. 9. Effect of ARAP3 overexpression on endogenous RhoA, Rac1 and Cdc42. Levels of active GTP-bound RhoA, Rac1 and Cdc42 in the indicated Flp-In^TM 293 inducible cell lines, treated (+) or not treated (-) with 2 µg/ml doxycycline (DX) were determined by incubating cell lysate with GST fusion proteins containing the Rhotekin RBD (to detect RhoA GTP) or the PAK1 RBD (to detect Rac1 GTP and Cdc42 GTP). Affinity precipitates and samples of whole cell lysate (2% of that used in A.P.) were analysed by SDS-PAGE and western blotting (WB) with the indicated antibodies. The results shown are representative of two experiments.