Inhibition of TF expression by NF-κB and Erk-1/2 inhibitors.

(A) SMC cultures were incubated with the indicated inhibitors for 1 hour prior to stimulation with OSM (10 ng/mL) for a further hour. Total RNA was extracted at the end of each incubation and analyzed for TF mRNA expression. The blots were rehybridized with G3PDH to demonstrate equal RNA loading. (B) SMCs were preincubated for 1 hour with NF-κB and Erk-1/2 inhibitors followed by stimulation with OSM (10 ng/mL) for 10 minutes at 37°C. Extracted proteins (30 μg) were separated on a 4% to 12% Bis-tris gradient gel and transferred to a PVDF membrane. Phosphorylated and total Erk-1/2 were visualized by incubating the filters with a monoclonal antibody to p42/44 Erk-1/2 and a pan Erk-1/2 monoclonal antibody. Experiments were repeated twice.