Page 213 - Haematologica - Vol. 105 n. 6 - June 2020

Cotl1 regulates shear-dependent thrombus formation
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Figure 2. Cotl1 is required for thrombus formation and stability at high shear. (A-C) Assessment of platelet adhesion (A and B) and aggregate formation (A and C) on Horm collagen (70 μg mL-1) under flow (150-3 000 s-1) in heparinized whole blood of WT and Cotl1-/- mice. Values are mean±standard deviation (SD) (n=12). Scale bar, 50 μm. (D and E) Heparinized whole blood of WT and Cotl1-/- mice was perfused over a von Willebrand factor (vWF)-coated cover slip for 4 minutes (min) at a shear rate of 3,000 s-1. (D) Representative phase contrast images taken at the end of the perfusion time and (E) analysis of the number of adherent platelets per mm2 ±SD (left), as well as the rolling velocity calculated from the distance a platelet covered per minute in μm ±SD (right) was performed. Images were acquired with a Zeiss Axiovert 200 inverted microscope (40x/0.6 oil objective). Images are representative of at least 12 animals per group. Scale bar, 50 μm. Unpaired Student t-test: **P<0.01; *P<0.05. (F-H) Real-time deformability cytometry (RT-DC),27 a method for continuous mechanical characterization of cells which are deformed by deceleration at the stagnation point of fast extensional flow. (F) Scheme depicting the principle of real-time deformability cytometry (RT-DC.) (G) Representative dot plots showing the relative deformation, as well as the (H) the size of washed WT and Cotl1-/- platelets. Values are mean±SD (n=3). *P<0.1; **P<0.01. A.U. : arbitrary units.
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