Figure 3.
Potent thrombolytic and antiplatelet activity of caADAMTS13 following distal thrombotic MCAo in mice. (A) The extent of VWF and fibrin(ogen) deposition at the site of the MCAo (#) and in the downstream vessels (white arrows) was visualized by immunofluorescence (IF). (B-C) Thresholded single-channel images were used to quantify the total area of VWF and fibrin(ogen)+ staining, which was normalized for background fluorescence using an equivalent area of the contralateral hemisphere. (D) The number of CD41+ particles, with an area greater than 0.2 μm2, was measured using automated particle counting applied to thresholded images in ImageJ. (E) Platelet aggregates (positive for both VWF and CD41) in the penetrating arterioles are indicated by white arrows. (F-G) The extent of ET-1 staining in the pial vessels was visualized by IF (white arrows) and was quantified/normalized using the same procedure. All images are a representative field of view, chosen to visualize the main site of thrombosis, isolated from whole-brain imaging performed using Slidescanner. Scale bars, 100 μm. Error bars represent mean ± standard deviation. Single comparisons were performed using an unpaired Student t test. *P < .05, **P < .01.

Potent thrombolytic and antiplatelet activity of caADAMTS13 following distal thrombotic MCAo in mice. (A) The extent of VWF and fibrin(ogen) deposition at the site of the MCAo (#) and in the downstream vessels (white arrows) was visualized by immunofluorescence (IF). (B-C) Thresholded single-channel images were used to quantify the total area of VWF and fibrin(ogen)+ staining, which was normalized for background fluorescence using an equivalent area of the contralateral hemisphere. (D) The number of CD41+ particles, with an area greater than 0.2 μm2, was measured using automated particle counting applied to thresholded images in ImageJ. (E) Platelet aggregates (positive for both VWF and CD41) in the penetrating arterioles are indicated by white arrows. (F-G) The extent of ET-1 staining in the pial vessels was visualized by IF (white arrows) and was quantified/normalized using the same procedure. All images are a representative field of view, chosen to visualize the main site of thrombosis, isolated from whole-brain imaging performed using Slidescanner. Scale bars, 100 μm. Error bars represent mean ± standard deviation. Single comparisons were performed using an unpaired Student t test. *P < .05, **P < .01.

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