Fig. 6. Rho inhibition reverts the phenotype induced by TTC3 overexpression in PC12 cells. (A) PC12 cells were transfected with empty GFP or GFP-TTC3 constructs, either alone or in combination with HA-C3 expression plasmid. Cultures were treated with NGF after 12 hours, allowed to differentiate for 3 days and processed for IF. (B) Quantitative analysis of differentiated cells in the above experiment, performed as in Fig. 2. (C) HEK293 cells (upper panel) or PC12 cells (lower panel) were transfected with empty GFP or GFP-TTC3 expression vectors and analyzed after 48 hours by Rho pull-down assay, using recombinant Rhotekin Rho-binding domain (GST-RBD).