Colonies of MDCK cells in 0.2% FCS were microinjected with a plasmid encoding PAK4wt at a concentration of 30 ng/μl or PAK4ΔGBD at a concentration of 100 ng/μl and incubated for 3 hours followed by a further 2.5 hours incubation in the absence or presence of HGF. LY294002 was added 2.5 hours post injection for 30 minutes before a further incubation for 2.5 hours in the absence or presence of HGF. Cells were then fixed and stained for PAK4 expression and F-actin. Using confocal microscopy, cells were then scored for the localisation of PAK4 proteins. A total of 60-100 cells were counted over two experiments.