Fig. 9. Removal of cholesterol affects the TPA-induced localization of ARF6 with Rac1 at the free surface of the cells. Serum-starved cells were untreated or incubated with or without 5 mM mßCD for 30 minutes at 37°C prior to addition of 0.1 µM TPA. After 10 minutes at 37°C the cells were fixed, permeabilized and stained with antibodies to visualize endogenous Rac1 and ARF6. Arrows indicate the localization of both Rac1 and ARF6 to the free surface where ruffling and macropinocytosis take place.