Effect of various intracellular signaling inhibitors on FVIIa and thrombin-induced TF mobilization. Fibroblasts were preincubated with inhibitors for 1 hour prior to the addition of FVIIa (A) or thrombin (B) (10 nM). Following FVIIa or thrombin treatment for 2 hours at 37°C, the cells were fixed, permeabilized, and immunostained for TF and the Golgi. Number of total cells and cells with intense TF staining in the Golgi were counted in multiple fields of 3 to 4 independent experiments, and the data were shown as percentage of cells with TF in the Golgi. Presence of intense TF staining in the perinuclear region (C, arrow mark) was taken as TF in the Golgi, and the absence of visible perinuclear staining was taken as no TF in the Golgi. In most of the observed cells, TF staining in perinuclear region was either intense or unnoticeable. The concentration of the inhibitors used are as follows: LY29042, 10 μM; wortmannin, 0.1 μM; PD98059, 50 μM; SB203580, 25 μM; PP2, 10 μM; PP3, 10 μM; and Go6976, 1.0 μM. * and # denote that these values differ from the values obtained with no inhibitor but treated with FVIIa with a P value of < .05 and < .01, respectively.