Figure 2.
Figure 2. aHA mice are not protected against tail bleeding. (A) Tail bleeding in WT C57Bl/6J mice at different time points after injection of anti-FVIII antibody (0.25 mg/kg; n = 4-5). Bleeding was determined by measuring the mouse weight-normalized blood loss (µL/g) after tail resection for 20 minutes. (B) Tail bleeding in WT C57Bl/6J mice (n = 5-10; filled circles) measured 2 hours after injection of different doses of anti-FVIII antibody (0-1 mg/kg). Tail bleeding in FVIII−/− BALB/c mice (n = 8; open circle) is shown as reference. (C) Bleeding determined by Hct at 24 hours after tail resection in WT C57Bl/6J mice (n = 5-10; filled circles) injected with different doses of anti-FVIII antibody (0-1 mg/kg). Hct at 24 hours in FVIII−/− BALB/c mice (n = 8; open circle) after tail resection is shown as reference. Representative thrombin-generation profiles (D), endogenous thrombin potential (ETP; nM × minutes) (E), peak height (nM) (F), and time to peak (TTP; minutes) (G) of plasma samples taken 24 hours after injection of anti-FVIII antibody (0-1 mg/kg) in WT BALB/c mice (n = 3-4 per antibody dosage) or FVIII−/− BALB/c mice (n = 3). (H) Representative uPA-mediated plasma clot lysis profiles from a FVIII−/− BALB/c mouse (red) or WT BALB/c mouse injected 24 hours before with either saline (blue) or anti-FVIII antibody (0.25 mg/kg; green). The 50% clot lysis time (CLT50%), representing the time between maximal clot formation and half-maximal clot lysis, is indicated by arrows. (I) CLT50% (minutes) of plasma samples taken 24 hours after injection of anti-FVIII antibody (0-1 mg/kg) in WT BALB/c mice (n = 3-4 per antibody dosage) or FVIII−/− BALB/c mice (n = 3). Thrombin-generation and clot lysis data are the mean of 2 independent experiments for each sample. Kruskal-Wallis test followed by Dunn’s post hoc multiple comparison test (A-C) or 1-way analysis of variance followed by Dunnett’s post hoc multiple comparison test (D-I) was used to compare experimental mice with WT mice injected with saline. *P < .05, **P < .01, ***P < .001, ****P < .0001. KO, knockout; ns, not significant; OD, optical density.

aHA mice are not protected against tail bleeding. (A) Tail bleeding in WT C57Bl/6J mice at different time points after injection of anti-FVIII antibody (0.25 mg/kg; n = 4-5). Bleeding was determined by measuring the mouse weight-normalized blood loss (µL/g) after tail resection for 20 minutes. (B) Tail bleeding in WT C57Bl/6J mice (n = 5-10; filled circles) measured 2 hours after injection of different doses of anti-FVIII antibody (0-1 mg/kg). Tail bleeding in FVIII−/− BALB/c mice (n = 8; open circle) is shown as reference. (C) Bleeding determined by Hct at 24 hours after tail resection in WT C57Bl/6J mice (n = 5-10; filled circles) injected with different doses of anti-FVIII antibody (0-1 mg/kg). Hct at 24 hours in FVIII−/− BALB/c mice (n = 8; open circle) after tail resection is shown as reference. Representative thrombin-generation profiles (D), endogenous thrombin potential (ETP; nM × minutes) (E), peak height (nM) (F), and time to peak (TTP; minutes) (G) of plasma samples taken 24 hours after injection of anti-FVIII antibody (0-1 mg/kg) in WT BALB/c mice (n = 3-4 per antibody dosage) or FVIII−/− BALB/c mice (n = 3). (H) Representative uPA-mediated plasma clot lysis profiles from a FVIII−/− BALB/c mouse (red) or WT BALB/c mouse injected 24 hours before with either saline (blue) or anti-FVIII antibody (0.25 mg/kg; green). The 50% clot lysis time (CLT50%), representing the time between maximal clot formation and half-maximal clot lysis, is indicated by arrows. (I) CLT50% (minutes) of plasma samples taken 24 hours after injection of anti-FVIII antibody (0-1 mg/kg) in WT BALB/c mice (n = 3-4 per antibody dosage) or FVIII−/− BALB/c mice (n = 3). Thrombin-generation and clot lysis data are the mean of 2 independent experiments for each sample. Kruskal-Wallis test followed by Dunn’s post hoc multiple comparison test (A-C) or 1-way analysis of variance followed by Dunnett’s post hoc multiple comparison test (D-I) was used to compare experimental mice with WT mice injected with saline. *P < .05, **P < .01, ***P < .001, ****P < .0001. KO, knockout; ns, not significant; OD, optical density.

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