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Fig. 6. NIH-3T3 cells were co-transfected with GEF-H1 and PAK4. Expression of GEF-H1 and PAK4 was detected by fluorescence imaging of EGFP for GEF-H1 and PAK4 antiserum (#933) followed with anti-rabbit fluorescent rhodamine for PAK4. Fluorescent coumarin-phallicidin was used to examine the actin cytoskeleton. Anti-ß-tubulin followed by anti-mouse Marina Blue conjugate was used to stain for the MT cytoskeleton. (A) GEF-H1M together with PAK4 and the mutant PAK4-S474E. Arrow a highlights an actin halo seen with overexpressed GEF-H1M; arrow b indicates stress fibers organized with actin cables; arrow c highlights actin-rich lamellipodia in cells co-expressing GEF-H1M and the mutant PAK4-S474E. (B) GEF-H1M together with vector and the mutants PAK4-S474E and PAK4 K350,351A. Note the non-polarized partial dissolution of the MT cytoskeleton in the presence of the mutant PAK4-S474E. Arrow d indicates lack of MTOC. Bar, 10 µm.





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