Fig. 1. CNF-1 activates RhoA in T84 intestinal epithelial cells. Confluent T84 intestinal epithelial monolayers were incubated with either CNF-1 (1 nM) or vehicle for 24 hours, and RhoA, Rac1 and Cdc42 activation status was examined by pull-down assays involving binding to the GTPase-binding domains of rhotekin or PAK-1 conjugated to agarose beads. Total Rho, Rac and Cdc42 protein levels were similar in lysates from control and CNF-treated monolayers. However, in pull-down activation assays, more Rho, Rac and Cdc42 were recovered from CNF-treated cells relative to controls, indicating increased activation of these GTPases.