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. 2002 Jul;22(13):4587–4597. doi: 10.1128/MCB.22.13.4587-4597.2002

FIG. 6.

FIG. 6.

Cell attachment to FN activates a Cdc42/Rac1 signaling pathway. (A) ECV 304 cells transiently transfected with dn N17Cdc42 or dn N17Rac1 cDNA or mock transfected were plated on FN for 20 min. Cell lysates were analyzed for active GTPases by binding to a GST-PAK-CRIB domain fusion protein, followed by Western blotting with anti-Rac1 or anti-Cdc42 antibodies. Total levels of Rac1, Cdc42, and actin were analyzed by Western blotting of original cell lysates. (B) ECV 304 cells transfected with Cdc42 or dn Rac1 cDNA or mock transfected were allowed to attach to FN for the indicated times, and then activated FAK and ERK1/2 were analyzed by Western blotting with anti-phospho-FAKY397 PAb or anti-phospho-ERK1/2 PAb. The levels of c-myc were detected with anti-c-Myc MAb 9E10. Actin levels were analyzed as a loading control.