Fig. 3: Therapeutic treatment with DMF and 4-OI suppresses thrombin generation in vivo induced by LPS, E. coli, and S. aureus, improving survival in mice.

a–f Mice were intraperitoneally injected with 1 mg/kg LPS (2 h) followed by treatment with PBS, a 50 mg/kg DMF, b 50 mg/kg 4-OI, c 50 mg/kg baricitinib, or d 15 mg/kg 1H1 anti-TF antibody (for a further 4 h) via intraperitoneal injection. Citrated plasma was harvested and thrombin generation was assessed (n = 4 per group). Thrombin generation in a–d is compared relative to PBS-treated mice ±LPS challenge. e Peak thrombin in mouse citrated plasma treated as in a–d. PBS–LPS + PBS, P = 0.00000637; LPS + PBS–LPS + DMF, P = 0.0000008; LPS + PBS–LPS + 4-OI, P = 0.0000167; LPS + PBS–LPS + 1H1, P = 0.0000000055. f Total thrombin generation (ETP = endogenous thrombin potential) in mouse citrated plasma treated as in a–d. LPS + PBS–LPS + 1H1, P = 0.00000135. g Kaplan–Meier survival curve of mice injected intraperitoneally with 15 mg/kg LPS (24 h) followed by treatment with PBS, 50 mg/kg DMF, 50 mg/kg 4-OI, or 200 IU/kg heparin (for a further 48 h) (n = 10 per group). LPS + PBS–treatment groups, P = 0.0000003. h Mice from g were scored clinically (assessing weight loss, activity level, eye closure, and appearance of fur and posture) every 6 h for the duration of the 72 h experiment. LPS + PBS–LPS + DMF, P = 0.00000001997. Data points indicate individual mice in g, h. For g, h Mantel–Cox survival analysis was performed. i Mice were intraperitoneally injected with 1 × 10⁷ CFU E. coli (CFT073) and co-treated with PBS, 50 mg/kg DMF, or 50 mg/kg 4-OI (6 h), followed by supplemental treatment via intraperitoneal injection with PBS, 50 mg/kg DMF, or 50 mg/kg 4-OI (for a further 18 h). Citrated plasma was harvested and thrombin generation was assessed (n = 5 per group). j Peak and k total thrombin generation in mouse citrated plasma treated as in i (n = 5 per group). l BMDMs were pre-treated with DMSO, DMF, or 4-OI (1 h) before infection with 5 × 10⁶ CFU S. aureus (USA300-LAC) (3 h) and harvesting cell lysates. F3 mRNA was quantified by qRT-PCR. m Mice were intraperitoneally injected with 5 × 10⁸ CFU S. aureus (PS80) and co-treated with PBS, 50 mg/kg DMF, or 50 mg/kg 4-OI (6 h), followed by supplemental treatment via intraperitoneal injection with PBS, 50 mg/kg DMF, or 50 mg/kg 4-OI (for a further 18 h). Citrated plasma was harvested and thrombin generation was assessed (n = 5 per group). n Peak thrombin generation in mouse citrated plasma treated as in m. o TF-positive regions (red) in the lungs of mice treated as in m. Representative lung tissue sections are shown. Magnification ×20. Scale bar = 10 μm. Data from a–k and m, n are mean ± SD. Data from l are mean ± SEM from three independent in vitro experiments. P values were calculated using a two-tailed Student’s t test for paired comparisons or one-way ANOVA for multiple comparisons. Source data are provided as a Source Data file.