Neutrophils contain TF in vitro and expressed it in vivo at the site of laser-induced injury. (A) In vitro detection of TF in neutrophils; Western blot analysis of protein extracts from purified Panc02 mouse pancreatic cancer cells, bone marrow neutrophils, circulating neutrophils, and washed platelets with 2 anti–mouse TF antibodies (left panel). Immunofluorescence microscopy of neutrophils (PMNs) isolated from mice with an antibody directed against mouse TF with an Alexa 488–conjugated secondary antibody. Negative control was performed with an irrelevant antibody and the secondary antibody. Original magnification, ×200; n = 3 (right panel). (B) Representative pictures showing the visualization of TF in the bloodstream of wild-type mice at the site of laser-induced injury in presence or absence of ICAM-1–blocking antibody using the mouse anti-TF antibody 1H1. TF is shown in yellow. Graph represents the sum of the medians of TF-integrated fluorescence intensity in wild-type mice in presence (32 thrombi in 3 mice) or absence (31 thrombi in 3 mice) of ICAM-1–blocking antibody. (C) Fibrin generation and thrombus formation at the site of laser-dye injury in low TF mice before and after infusion of isolated neutrophils (PMNs). Fibrin is depicted in green; platelets are in red. (D) Graph depicts medians of fibrin and platelet-integrated fluorescence intensity in low TF mice (32 thrombi, 3 mice) and low TF mice after infusion of PMNs (31 thrombi, 3 mice).