Fig. 7. Abrogation of Tc10- and RhoT-induced neurite extension in PC12 cells by dominant-negative mutants of N-WASP. (A,B) PC12 cells were cotransfected with the cDNA of Cdc42(G12V), Tc10(G18V) or RhoT(G30V) in pEF-BOS/Myc vector and that of N-WASP(H208D) (A) or N-WASPcof (B) in pcDL-SR vector. Forty-eight hours after the transfection, the cells were doubly stained with Myc1-9E10 to detect the expression of Cdc42(G12V) (a), Tc10(G18V) (c) and RhoT(G30V) (e) and with anti-N-WASP pAb (b,d,f). Note that the cells coexpressing the dominant-negative mutants of N-WASP with the small GTPases, indicated by arrows, do not extend neurites. Bar, 20 µm. (C) Ratio of the neurite-extending PC12 cells treated as above. The degree of neurite extension is expressed by multiples of the cell body diameter as indicated in the legend. More than 100 cells were counted in each experiment. The values are the means±s.d. of triplicate experiments.