Fig. 6. MCP-1 transiently activates Rho in mouse BMEC. (A) Confluent mBMEC were treated with 100 nM murine MCP-1 for 10, 20, 30 minutes, 1 or 2 hours. Cell lysates were subject to affinity precipitation using Rhotek in recombinant protein agarose conjugated which specifically precipitates active RhoA (Rho-GTP). Total Rho indicates total amount of active and inactive Rho in the mBMEC. The immunoblot represents one of three independent experiments. (B) Densitometric analysis of MCP-1-induced activation of RhoA. Data are means±s.e.m., n=3 independent experiments ^*P<0.001; (C) Confluent mBMEC were subject to treatment with a Rho kinase inhibitor Y27632 10 µM for 30 minutes) or with specific inhibitor of RhoA, C3 exoenzyme (5 µg/ml for 18 hours), or transiently transfected with dominant negative mutant T19NRho. Western blot was performed using an antibody specific for RhoA.

Return to article