Depletion of neutrophils in mice decreases HRPII-mediated inflammation but not the procoagulant response. Mice were administered either control antibody or anti-Ly6G antibody (20 µg/g) 48 hours before HRPII treatment (10 µg/g). After 3.5 hours of HRPII treatment, mice were euthanized and perfused with PBS, and blood and organs were collected. The lower right lobe of the lung was lysed in tissue lysis buffer. (A) Lung tissue lysates were immunoblotted for VCAM1, ICAM1, MPO, fibrin(ogen), and β-actin. Densitometric analysis of expression of these proteins is presented in (B), (C), (D), and (E). β-actin was used as loading control. (F) Plasma level of the TAT complex was measured with established ELISA. (G) Plasma level of VWF was measured by ELISA using a commercial kit. (H) Blood platelet counts were determined using a veterinary hematology analyzer. (I) To analyze the effect of neutrophil depletion on lung vascular permeability, 1% Evans blue dye was injected 3 hours after HRPII treatment. After 30 minutes, animals were euthanized and perfused with PBS, lung tissue samples were collected, and vascular permeability was measured from the amount of Evans blue dye leaked into the lung as described in “Materials and methods.”