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Fig. 1. Effect of Rac2^wt and Rac2^D57N expression in bone-marrow-derived macrophages on the actin cytoskeleton and the microtubule networks. Macrophages plated on coverslips were transfected with expression constructs for Rac2^wt in pIRES2-EGFP (A-D,G), Rac2^D57N in pIRES2-EGFP (E,F), empty vector pIRES2-EGFP (H) or FLAG-Rac2^D57N in pCDNA3.1 (I-L). Cells were incubated for 24 hours in macrophage medium and treated as described in Materials and Methods. F-actin was stained with rhodamine-phalloidin (A,C-E,J,L). Microtubules were immunostained with an antibody to -tubulin (F-H). Transfected cells were identified by EGFP expression (A-H) or by immunostaining for FLAG (I-L). Arrowheads indicate transfected cells. Arrows indicate colocalization of Flag-Rac2^D57N and F-actin at the cell membrane. Asterisks indicate colocalization of Rac2^D57N with aggregates of F-actin. Scale bar, 10 µm.

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