Fig. 6. ARAP3-mediated membrane process formation is Rho GAP dependent, blocked by activated RhoA expression, and enhanced by mutation of the tyrosine phosphorylation sites Y1399 and Y1404. (A) The indicated Flp-In^TM T-Rex^TM 293 cell lines were grown overnight on laminin- or fibronectin-coated dishes in the presence (+) or absence (-) of 2 µg/ml doxycycline (Dox). Multiple random fields of cells were imaged as in Fig. 5 and the proportion of cells with process(es) >1 cell diameter was scored. Between 200 and 350 cells were scored per sample. Data are means ± s.e.m. from duplicate determinations in two independent experiments. (B) Cell lines treated with doxycycline overnight to induce ARAP3 proteins were transfected with plasmids encoding green fluorescent protein (GFP) plus a 10-fold excess of RhoA V14 or empty vector (Vector). Live transfected cells expressing GFP were visualised 24 hours post-transfection by confocal fluorescence microscopy using a water immersion 60x objective.