Colocalization analysis of Toca-1 and N-WASP expressing cells.

Cells were transfected with Toca-1 and N-WASP and left for 24–36 hr. After fixation cells were incubated with primary antibody followed by secondary antibody for markers of membrane trafficking pathways as described in the Material and methods section. PH domains were visualized with GFP, or CFP. Olympus FV1000 confocal microscope software was used for colocalization analysis. Where Pearson colocalization coefficient (CC) of; <0.25 indicates no colocalization (-), 0.25–0.5 indicates potential colocalization (*), 0.5–0.7 indicates some colocalization (**) and >0.7, indicates high colocalization (***). The CC is an average +/− S. D., with n = 8–10, from 3 experiments.