Extended Data Fig. 3. Absence of GPV restores thrombotic and haemostatic defects in the absence of GPVI.

GPVI was depleted from the platelet surface by injection of the anti-GPVI mAb JAQ1. Confirmation of GPVI depletion by Western blot analysis (a) and flow cytometry (b); representative of 3 independent experiments. (c) Quantification and representative images (scale bar: 50 μm) (d) of thrombus formation upon FeCl₃-induced injury of mesenteric arterioles. Thrombus formation in no more than two arterioles of each mouse were analysed; data points represent measurements of one arteriole. WT: 18 arterioles of 11 mice, Gp5^−/−:12 arterioles of 8 mice, WT + JAQ1: 15 arterioles of 8 mice, Gp5^−/−+ JAQ1: 12 arterioles of 7 mice. Two-tailed Fisher’s exact test for open vs. occluded vessels, p=0.0005 (WT vs. WT +JAQ1), p=0.0031 (Gp5^−/− vs. WT + JAQ1; Gp5^−/− + JAQ1 vs. WT+JAQ1); one-way ANOVA followed by Tukey’s test for multiple comparisons to compare occluded vessels, ## p= 0.0034 (WT vs. Gp5^−/−), # p=0.0401 (WT vs. Gp5^−/− + JAQ1) compared to JAQ1-treated WT mice (p=0.0031). * compared to JAQ1-treated WT mice. (e) The abdominal aorta was mechanically injured by a single firm compression with a forceps and blood flow was monitored with a Doppler flowmeter. Time to cessation of blood flow is shown. Each symbol represents one mouse. Two-tailed Fisher’s exact test for open vs. occluded vessels, one-way ANOVA followed by Dunn’s test for multiple comparisons to compare occluded vessels. WT: n=18, Gp5^−/−: n=16, WT + JAQ1: n=7, Gp5^−/−+ JAQ1: n=7, ***WT +JAQ1 vs. WT and vs. Gp5^−/−: p<0.0001, ** WT +JAQ1 vs. Gp5^−/−+JAQ1: p=0.0047), ### compared to untreated WT mice (WT+JAQ1: p=0.0003). (f) Haemostatic function was assessed using a tail bleeding assay on filter paper. Each symbol represents one mouse. WT: n=18, Gp5^−/−: n=17, WT + JAQ1: n=12, Gp5^−/−+ JAQ1: n=9. one-way ANOVA followed by Dunn’s test for multiple comparisons to compare occluded vessels, p=0.0001’*, ^# p < 0.05; **, ^## p < 0.01; ***, ^### p < 0.001.