Figure 4
Figure 4. Mice expressing murine VWF/p.S1494C-p.A1534C are thrombocytopenic and present with schistocytes. Every mouse used in this study is VWF−/−. (A) Multimer analysis of plasma taken from mice expressing mVWF/p.S1494C-p.A1534C (mice 1 and 2) or mVWF/WT at day 4 and day 7 after hydrodynamic injection using 2% SDS-agarose gel electrophoresis. Gels were loaded with 0.1 μg antigen per lane. All multimers originate from the same multimer gel. (B) Coulter platelet counts and plasma VWF antigen ratio were measured at days 4 and 7 after injection. Only mice with VWF antigen levels within the 300% to 1000% normal range were included in this study. Significant differences were found in platelet counts between mVWF/WT-expressing mice and mVWF/p.S1494C-p.A1534C-expressing mice. Although mVWF/WT mice have normal platelet counts at both time points, 5 of 7 mVWF/p.S1494C-p.A1534C mice were found to be thrombocytopenic at day 4 and 6 of 7 by day 7. (C) Blood smears were prepared with heparinized blood from mice injected with normal and mutant pLIVE-mVWF constructs and subjected to GIEMSA staining. Although mice expressing mVWF/WT do not present with any anomalies, mice expressing mVWF/p.S1494C-p.A1534C present with occasional platelet aggregates (black arrow), schistocytes (white arrows), and fewer platelets and erythrocytes. For each mouse, 2 blood smears were prepared, and 5 mice were analyzed for each construct. Wide field images were captured at an original magnification of ×63 (1.4 aperture) and high-magnification images at ×100 (1.4 aperture). All images were captured on a LSM 510 Meta confocal microscope at room temperature. All images were captured and processed with the LSM 510 software.

Mice expressing murine VWF/p.S1494C-p.A1534C are thrombocytopenic and present with schistocytes. Every mouse used in this study is VWF−/−. (A) Multimer analysis of plasma taken from mice expressing mVWF/p.S1494C-p.A1534C (mice 1 and 2) or mVWF/WT at day 4 and day 7 after hydrodynamic injection using 2% SDS-agarose gel electrophoresis. Gels were loaded with 0.1 μg antigen per lane. All multimers originate from the same multimer gel. (B) Coulter platelet counts and plasma VWF antigen ratio were measured at days 4 and 7 after injection. Only mice with VWF antigen levels within the 300% to 1000% normal range were included in this study. Significant differences were found in platelet counts between mVWF/WT-expressing mice and mVWF/p.S1494C-p.A1534C-expressing mice. Although mVWF/WT mice have normal platelet counts at both time points, 5 of 7 mVWF/p.S1494C-p.A1534C mice were found to be thrombocytopenic at day 4 and 6 of 7 by day 7. (C) Blood smears were prepared with heparinized blood from mice injected with normal and mutant pLIVE-mVWF constructs and subjected to GIEMSA staining. Although mice expressing mVWF/WT do not present with any anomalies, mice expressing mVWF/p.S1494C-p.A1534C present with occasional platelet aggregates (black arrow), schistocytes (white arrows), and fewer platelets and erythrocytes. For each mouse, 2 blood smears were prepared, and 5 mice were analyzed for each construct. Wide field images were captured at an original magnification of ×63 (1.4 aperture) and high-magnification images at ×100 (1.4 aperture). All images were captured on a LSM 510 Meta confocal microscope at room temperature. All images were captured and processed with the LSM 510 software.

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