Fig. 5. Colocalization of wt and mutant Nod2 proteins with endogenous Rac1. (A) Rac1 colocalization with F-actin in membrane ruffles. COS-7 cells were stained for Rac1 with the anti-Rac1 mAb and for F-actin with TRITC-phalloidin. Images were obtained by confocal microscopy (Leica TCS NT). (B) COS-7 cells were transfected with the following amounts of plasmid encoding the HA-tagged or FLAG-tagged wt or mutant Nod2 proteins: pcDNA₃-HA-wt Nod2 and pcDNA₃-HA-CARDs (1 µg each), pcDNA₃-HA-LRRs (4 µg), pcDNA₃-HA-CARDs (0.5 µg), pcDNA3-HA-LRRs (2 µg), pcDNA₃-FLAG-NOD (2 µg). In each case, the total DNA amount was adjusted to 4 µg with pcDNA3. Twenty-four hours post transfection, cells were stained for the wt or mutant Nod2 proteins with monoclonal anti-HA or rabbit anti-FLAG antibodies and for endogenous Rac1 with a monoclonal antibody. Images were obtained by confocal microscopy (Leica TCS NT). Arrows indicate areas of colocalization.