PDI antibodies or exogenously added PDI failed to alter TF-FVIIa coagulant and cell signaling activities. (A). MDA-MB-231 cells were incubated with control IgG, anti-PDI IgG (100 μg/mL), or anti-TF IgG (50 μg/mL) for 30 minutes and then treated with control vehicle or HgCl2 (25 μM) for 15 minutes. TF-FVIIa coagulant activity was determined as described in Figure 1 legend. (B) MDA-MB-231 cells were incubated with control vehicle or various concentrations of recombinant PDI for 30 minutes before TF-FVIIa coagulant activity was measured. (C) MDA-MB-231 cells were incubated with control IgG, anti-PDI IgG, recombinant PDI (10 nM), or anti-TF IgG for 30 minutes and then treated with FVIIa (10 nM) for 1 hour. TF-FVIIa-induced IL-8 gene expression was determined by Northern blot analysis. p denotes polyclonal antibodies (100 μg/mL) and m, monoclonal antibodies (RL77 for PDI and 10H10 for TF, 10 μg/mL). (A,B) Data are expressed as mean plus or minus SEM (n = 3).
