Effect of dominant negative kinase mutants on the synthesis of MCP-1 by HUVEC. Cells were transfected in a 1:3 ratio with pGreenLantern expressing GFPS65T and either empty expression vector KRSPA or plasmids expressing dominant negative kinase mutants as indicated and stimulated with 2 ng/mL TNF- for 12 hours. Successfully transfected cells were analyzed for MCP-1 synthesis by flow cytometry as described in Materials and Methods. The data shown represent mean values ± SEM from four independent experiments. The asterisk indicates a statistically significant difference as compared with control (P < .05, Wilcoxon test).