FIGURE 4:

PI(3,4,5)P3 concentrates at nonopsonized-particle phagosomes but not in the presence of nocodazole. Cells were transiently transfected with AKT-PH-GFP to detect accumulation of PI(3,4,5)P3 at particle phagosomes. When exposed to (A) Ab-opsonized spherical silica or (B) ovalbumin-coated spherical silica, AKT-PH-GFP quickly concentrates at the site of the particle and dissociates once a particle has been internalized. (C) When cells are treated with 800 nM nocodazole, AKT-PH-GFP does not accumulate around nonopsonized- particle phagosomes. These data are representative of localization visualized on six separate days. (D) The rate of AKT-PH-GFP association with and dissociation from particle phagosomes. N = 12. Time zero represents maximum localization after particle–cell interaction, and error bars represent SEM.