File:A-Phosphatidylinositol-3-Kinase-Dependent-Signal-Transition-Regulates-ARF1-and-ARF6-during-Fcγ-pbio.0040162.sv004.ogv

English: Movie of ARF6 Activation in the Presence of LY294002 RAW264.7 macrophages co-transfected with ARF6-CFP and YFP-NGAT were incubated in 50 μM LY294002 for 30 min before the addition of IgG-opsonized erythrocytes. The presented images, left to right, are: phase contrast, R M and E D time-lapse images. Following binding of a particle at the right, ARF6-CFP was activated and pseudopod extension commenced. GTP-bound ARF6-CFP continued to accumulate in the phagocytic cup and was not deactivated during observation. The accumulation of ARF6-CFP at the arrested phagocytic cup can be seen in the R M image series (the blue band at the phagocytic cup represents a molar excess of ARF6-CFP relative to YFP-NGAT). The generation of ARF6-GTP in the absence of PI-3K function indicates ARF6 is activated at the phagosome by a PI-3K-independent GEF activity.