Increased VWF staining of the hepatic endothelium of <i>F8</i>^<i>-/y</i> mice.

<p>(<b>A</b>) Representative hepatic tissue sections of <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice stained with anti-human VWF antibody (100x and 400x magnification). (<b>B</b>) Visual scoring of the VWF stained hepatic tissue sections of <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 7,6). (<b>C</b>) Hepatic VWF transcript levels in <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 4,7). All data were expressed as means ± SEM. Statistical comparisons were performed using the Student’s <i>t</i>-test, ** p<0.01.</p

Mice deficient in the anti-haemophilic coagulation factor VIII show increased von Willebrand factor plasma levels - Table 1

<p>Mice deficient in the anti-haemophilic coagulation factor VIII show increased von Willebrand factor plasma levels</p> - Table

Increased VWF levels in the plasma of <i>F8</i>^<i>-/y</i> mice.

<p>(<b>A</b>) VWF antigen levels in plasma of <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 9). (<b>B</b>) VWF multimer analysis of plasma samples from <i>F8</i>^<i>+/y</i> vs <i>F8</i>^<i>-/y</i> mice (n = 5). (<b>C</b>) ADAMTS13 levels in plasma of <i>F8</i>^<i>+/y</i> vs <i>F8</i>^<i>-/y</i> mice (n = 7). (<b>D</b>) VWF antigen levels in plasma of <i>F8</i>^<i>-/y</i> mice, 2h after tail vein injection of recombinant FVIII (Kogenate) at a dose of 1.5 U per 30g body weight or with 0.9% NaCl solution as a vehicle control (n = 11,10,12). All data were expressed as means ± SEM. Statistical comparisons were performed using the Student’s <i>t</i>-test or one-way ANOVA, * p< 0.05, ** p<0.01.</p

Bleeding phenotype of the <i>F8</i>^<i>-/y</i> mice.

<p>(<b>A</b>) Patella diameter before and 24h after joint bleeding induction to the right knee (n = 7,6) of C57BL/6J and <i>F8</i>^<i>-/y</i> with the left knee (n = 6,6) set as a control. (<b>B</b>) Representative rotational thromboelastometry (ROTEM) graphs of a C57BL/6J and a <i>F8</i>^<i>-/y</i>, illustrating clotting time (green), clot formation time (pink) and clot firmness (blue). (<b>C</b>) Clotting times and (<b>D</b>) clot formation times of C57BL/6J, <i>F8</i>^<i>-/y</i>, and <i>F8</i>^<i>-/y</i> blood samples reconstituted with 2.5 U/ml of human recombinant FVIII (n = 3). All data were expressed as means ± SEM. Statistical comparisons were performed using one-way ANOVA or two-way ANOVA * p< 0.05, ** p<0.01, ***p<0.001.</p

Low-grade inflammatory phenotype in the liver of <i>F8</i>^<i>-/y</i> mice.

<p>(<b>A</b>) FVIII (n = 5,4), (<b>B</b>) TNFα (n = 6,7), (<b>C</b>) CD45 (n = 7), and (<b>D</b>) TLR4 (n = 6,4) hepatic transcript levels of <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice. (<b>E</b>) Representative hepatic tissue sections of <i>F8</i>^<i>+/y</i> vs <i>F8</i>^<i>-/y</i> stained with anti-mouse F4/80 antibody (200x magnification). (<b>F</b>) Visual scoring of the F4/80 stained hepatic tissue sections (n = 5). (<b>G</b>) Hepatic transcript levels of the macrophage marker F4/80 in <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 4,7). (<b>H</b>) Hepatic SAA3 transcript levels in <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 5,5). (<b>I</b>) Serum glutamate-pyruvate-transaminase levels in <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 5,5). (<b>J</b>) Serum glutamat-oxalacetat-transaminase levels in <i>F8</i>^<i>+/y</i> (WT) vs <i>F8</i>^<i>-/y</i> mice (n = 6,5). All data were expressed as means ± SEM. Statistical comparisons were performed using the Student’s <i>t</i>-test, * p< 0.05, ***p<0.001, ****p<0.0001.</p

Impaired thrombin generation by platelets from <i>F8</i>^<i>-/y</i> mice.

<p>(<b>A</b>), (<b>B</b>) and (<b>C</b>) Representative thrombograms of a <i>F8</i>^<i>+/y</i> (WT) vs a <i>F8</i>^<i>-/y</i> mouse. Thrombin generation in unstimulated platelet-rich plasma (PRP) (<b>A</b>), in PRP induced by 1 pM tissue factor (TF) (<b>B</b>), and in PRP induced by 0.1 U/ml thrombin (thr) (<b>C</b>), respectively. (<b>D</b>) Thrombin peak in unstimulated PRP (n = 5,2), PRP plus 1 pM TF (n = 5,2), and 0.1 U/ml thr-stimulated PRP (n = 5,2). (<b>E</b>) Endogenous thrombin potential in unstimulated PRP (n = 5,2), PRP triggered by 1 pM TF (n = 5,2), and 0.1U/ml thr-stimulated platelets in PRP (n = 5,3), analyzed by calibrated automated thrombography of <i>F8</i>^<i>+/y</i> vs <i>F8</i>^<i>-/y</i> mice. All data were expressed as means ± SEM. Statistical comparisons were performed using the Student’s <i>t</i>-test, * p< 0.05, ** p<0.01, ***p<0.001.</p