Effect of FVIIa on TNF-α-induced MAPK signaling and NF-κB and AP-1 activation in endothelial cells. HUVEC, serum-deprived for 12 h, were treated with FVIIa (100 nM) for 1 hour and then with TNF-α (5 ng/mL) for 30 minutes in serum-free medium. The cell lysates were subjected to immunoblotting with specific antibodies to analyze activation of ERK1/2 (A), JNK1/2 (B), p38 MAPK (C), NF-κB (p65 phosphorylation; D) and AP-1 (c-Jun phosphorylation; E). p, phospho; t, total. Bar graphs below immunoblots depict the quantification of the phosphorylated signal normalized to the total protein by densitometric analysis of the bands using ImageJ software. Data represent the mean ± SEM of 3 independent experiments. *P < .05; **P < .01; ***P < .001.