Effect of poly I:C on clotting time, TF as well as TM at the activity, protein, and mRNA level in monocytes. (A) Poly I:C or poly dI:dC (50 μg/mL) and 100 ng/mL LPS were used to stimulate monocytes for 2 hours, and then mRNA was isolated for RT-PCR analysis. The expression levels of TF, TM, IFN-β, IL-6 and β-actin were determined by RT-PCR. (B) Monocytes were stimulated with 50 μg/mL poly I:C and 100 ng/mL LPS for 5 hours, and thereafter TF and TM activity was measured. Results are presented as relative activity; mean plus SEM of triplicate wells. (C) Monocytes were stimulated with various concentrations of 10 to 100 μg/mL poly I:C or 100 ng/mL LPS for 5 hours, and the protein levels for TF and TM were determined by Western blot analysis using the respective antibodies. (D) Nucleic acids (10 μg/mL) were used to stimulate monocytes for 5 hours, and then cell extracts were prepared, and plasma clotting time was determined. LPS (100 ng/mL) was used as a standard activator. Results are presented as mean plus SEM of triplicate wells as clotting time (in seconds).