Effect of thiol-oxidizing and -reducing agents on cell surface tissue factor activity of stimulated endothelial cells and fibroblasts. (A) Monolayers of HUVEC were stimulated with tumor necrosis factor α (TNF-α) (20 ng/mL) + IL-1β (20 ng/mL) for 6 hours at 37°C to induce TF expression. Stimulated HUVEC were treated with various concentrations of HgCl2, reduced glutathione, or DTT for 5 minutes, and the oxidizing/reducing agent was removed and cells were washed twice before adding FVIIa (10 nM) and factor X (175 nM) to measure cell surface TF activity as the rate of factor Xa generated in a chromogenic assay. (B) Same as panel A except fibroblasts were used in place of stimulated endothelial cells. Data are expressed as mean plus or minus SEM (n = 3).
