Ex vivo and in vivo thrombus formation in Ptpn11^T468M/+ mice. (A) Heparinized whole blood from Ptpn11^T468M/+ mice, where platelets were labeled with DiOC₆, was perfused through Cellix Vena8 Fluoro+ biochips at arterial physiological shear rates of 250 s⁻¹ and 500 s⁻¹ on fibrillar collagen matrix during 2 minutes. Thrombus formation was visualized in real time by videomicroscopy. Representative 2- and 3-dimensional images were respectively obtained after processing with ImageJ and Imaris software (scale bar, 20 μm). Surface covered by platelets and thrombus volume were quantified by using ImageJ software (mean ± SEM; n = 6 mice of each genotype). (B) Heparinized whole blood from Ptpn11^T468M/+ mice was treated or not with ARC69931MX (10 μM) and indomethacin (10 μM) over 30 minutes; platelets were labeled with DiOC₆ and perfused through Cellix Vena8 Fluoro+ biochips at arterial physiological shear rate of 500 s⁻¹ on fibrillar collagen matrix over 2 minutes. Representative 3-dimensional images were obtained after processing with Imaris software (scale bar, 20 μm) at 2 minutes of perfusion. At the end of the perfusion, thrombus volume was quantified by using ImageJ software (mean ± SEM; n = 6 mice of each genotype). (C) Time to blood flow cessation (occlusion time) was measured with a transonic flow probe at the carotid artery of mice after exposure to 7% FeCl₃ for 3 minutes. Each point represents a mouse. (D) Time required for the bleeding to stop after 3-mm tail-tip transection of anesthetized mice. Each point represents a mouse. *P < .05, **P < .01, ***P < .001 according to 2-way ANOVA and 1-sample t test.