TF cytoplasmic domain negatively regulates LPS-induced TF expression in BMMs. (A) TF mRNA expression, TF clotting activity, and TF-VIIa–dependent Xa generation of unstimulated or 6-hour LPS (1 μg/mL)–stimulated BMMs from the indicated genotypes. TF mRNA induction was quantified by real-time PCR (Applied Biosystems); TF procoagulant activity and Xa generation were determined after standard octyl-glucoside cell lysates. (B) TLR4 and CD14 expression in unstimulated BMMs from the indicated genotypes by flow cytometry. Dotted histograms represent isotype-matched control antibody and the filled histograms represent specific surface marker staining. (C) TF levels in BMMs from the indicated genotypes that were stimulated with and without IFNγ for 6 hours. (D) TF clotting activity measured after 3 hours of stimulation. (E) Dose response of IFNγ-induced TF expression after 6 hours of stimulation in comparison to LPS-treated controls. TF activity was determined in standard octyl-glucoside cell lysates. *denotes different from wild-type, P < .05, t test.