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Figure 6

Figure 6


Fig. 6. ARAP2 and the cytoskeleton. (A,B) Role of ArfGAP and RhoGAP domains in effects of ARAP2 on cytoskeleton. Effect of replacing endogenous ARAP2 with recombinant ARAP2 on stress fiber (A) and focal adhesion (B) formation. U118 cells were transfected with empty vector, Flag-ARAP2, Flag-[R728K]ARAP2, or Flag-[K1190]ARAP2 with (right panel) or without (left panel) ARAP2 siRNA. Three days after transfection, cells were seeded onto fibronectin-coated slides, fixed, and stained with polyclonal anti-ARAP2 Ab and either Rhodamine-phalloidin to visualize actin (A) or monoclonal anti-paxillin Ab to visualize focal adhesions (B). (C) Effect of [Q67L]Arf6 on stress fibers in U118 cells. Cells were transfected with either plasmids for the expression of wild type or [Q67L]Arf6-HA. After 24 hours the cells were replated on fibronectin-coated coverslips, incubated for 6 hours in serum-free medium and then stained for epitope-tagged Arf6 and actin. (D) Effect of ARAP2 expression level on stress fibers induced by [Q63L]RhoA. Cells were co-transfected with siRNA targeting ARAP2 and an expression plasmid for [Q63L]RhoA, fixed and stained for RhoA and polymerized actin. Bars, 10 µm.





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