TF expression in endothelial cells transduced with various concentrations (MOI/cell) of adenovirus-carrying wild-type TF or TF mutant. HUVECs were transduced with various concentrations (MOI/cell) of adenovirus encoding wild-type TF, TF_C186S, TF_C209S, TF_C186S/C209S, or control adenovirus (β-Gal). After culturing cells for 48 hours, (A) the cell lysates were harvested and subjected to immunoblot analysis; (B) intact monolayers were used to measure cell-surface TF procoagulant activity in FX activation assay by adding FVIIa (10nM) and FX (175nM) and measuring the amount of FXa generated in a chromogenic assay (n = 3-4; mean ± SEM).