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Fig. 9. (A) Arf6 and Rab8 co-operate in inducing cell protrusions. Co-expression studies of Rab8-Q67L and Arf6-wt, Rab8-Q67L and Arf6-T27N, and Rab8-Q67L and Arf6-Q67L in HT1080 cells. Confocal microscopy showed that Rab8-Q67L/Arf6-wt expressing cells contain cell extensions, with Arf6/Rab8-specific tubular structures entering ruffling lamella. Rab8-Q67L and Arf6-T27N colocalized on vesicular and tubular structures. Co-expression of Arf6-Q67L and Rab8-Q67L resulted in the formation of spherical cells, where Arf6-Q67L was localized on large vacuolar structures, and Rab8-Q67L in the perinuclear region. (B) Arf6 cross-talks with Rab8 in the formation of cell surface extensions. HT1080 cells were mock-transfected or transfected with a construct encoding Rab8-Q67L. Co-transfection was done with constructs expressing Arf6-wt/Rab8-Q67L, Arf6-T27N/Rab8-Q67L and Arf6-Q67L/Rab8-Q67L. Cells showing neurite-like extensions were calculated as extension-positive. A total of about 50 cells were counted per experimental condition. Values, given as percentage of cells exhibiting cell surface extension in each scoring category, from three separate experiments are shown as the mean ±s.e.m. Representative fields of these cells are seen in the lowest row of A. Bars, 10 µM, 20 µM (lower panel).
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