FIGURE 3.

TNF-α-mediated up-regulation of cell adhesion molecules on endothelial cells. A, the PAR-1-dependent effect of APC (20 nm, 4 h), thrombin (20 nm, 4 h), and TFLLRN (100 μm, 4 h) on the modulation of the cell adhesion molecules VCAM-1, ICAM-1, and E-selectin in response to TNF-α (10 ng/ml, 4 h) was monitored by ELISA after treating cells with either PC-S195A (50 nm, 30 min) or FX-S195A (175 nm, 30 min) in the absence and presence of the function-blocking anti-PAR-1 antibody (25 μg/ml). B, endothelial cells were activated with TNF-α (10 ng/ml, 4 h) after treating cells with APC (20 nm, 4 h), FXa (20 nm, 4 h), and thrombin (20 nm, 4 h) ± FX-S195A (175 nm, 30 min), and the adherence of neutrophils (panel B) or THP-1 cells (panel C) was monitored as described under “Experimental Procedures.” In the presence of MβCD (10 mm, 1 h) and function-blocking anti-PAR-1 anti-PAR-2 antibodies (25 μg/ml, 30 min), cells were first incubated with these reagents. All results are means ± S.D. of three different experiments.