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. 2016 Feb 24;6:21956. doi: 10.1038/srep21956

Figure 6. Effects of compounds 1-4 on the inactivation and production of thrombin and factor Xa.

Figure 6

(A) Inhibition of thrombin (Th) by compound 1 (white box), 2 (light gray box), 3 (dark gray box), and 4 (black box) was measured using a chromogenic assay, as described in the “Materials and Methods” section. (B) The inhibition of factor Xa (FXa) by each compound was also monitored using a chromogenic assay, as described in the “Materials and Methods” section. Argatroban (A) or rivaroxaban (B) was used as a positive control. (C) The HUVEC monolayer was pre-incubated with FVa (100 pM) and FXa (1 nM) for 10 min with the indicated concentrations of each compound. Prothrombin was added at a final concentration of 1 μM and prothrombin activation was determined after 30 min, as described in the “Materials and Methods” section. (D) HUVECs were pre-incubated with the indicated concentrations of each compound for 10 min. TNF-α (10 ng/mL for 6 h)-stimulated HUVECs were incubated with FVIIa (10 nM) and FX (175 nM) in the absence or presence of anti-TF IgG (25 μg/mL); the FXa production was determined as described in the “Materials and Methods” section. *p < 0.05 vs. 0.0001 μM each compound, or argatroban (A) or 0.0001 μM each compound, or 0.0001 nM rivaroxaban (B), DMSO (C) or TNF-α alone (D).