Labeled with alexa fluor 647 protein labeling kit

Antibodies were added to sterile 0.9% NaCl solution (B. Braun Melsungen, Melsungen, Germany) to a final infusion volume of 100 μL. The used antibodies and antibody concentrations were: sinusoidal endothelial cells: rat anti-mouse CD31-PE, 10 μL of 200 μg/mL (cat.#12-0311-83, clone 390, eBioscience, San Diego, CA) or rat anti-mouse CD31-Alexa Fluor 647, 5 μL of 1000 μg/mL (cat.#16-0311-85, clone 390, eBioscience, labeled with Alexa Fluor 647 protein labeling kit, cat.#A-20173, Life Technologies, Carlsbad, CA); resting platelets: hamster anti-mouse CD49b-Alexa Fluor 647, 7 μL of 500 μg/mL (cat.#103511, clone HMα2, Biolegend, San Diego, CA); activated platelets, rat anti-mouse CD62P-FITC, 10 μL of 500 μg/mL (cat.#553744, clone RB40.34, BD Pharmingen, Franklin Lakes, NJ); neutrophils: rat anti-mouse Ly-6G (Gr-1)-FITC, 10 μL of 500 μg/mL (cat.#108406, clone R86-8C5, BioLegend). The mixture was infused into the penile vein directly before surgery using a 1 mL insulin syringe, after which the puncture wound was sealed with an electro-surgical cauterizer. Before the liver I/R experiments, in vivo thrombus staining by the CD62P-FITC antibodies was verified in a puncture-induced thrombosis model in the murine saphenous artery (N = 2, Fig. S2).