Fig. 2. The mAb GoH3 against 
6 integrin and the NE inhibitor ONO-5046 suppress leukocyte transmigration through IL-1ß-stimulated cremasteric venules in vivo. (A,B) WT or NE KO mice were injected i.v. with saline, control IgG or mAb GoH3 (both at 3 mg kg–1) 15 minutes before i.s. injection of saline or IL-1ß (30 ng). 2 hours later, the jugular vein was cannulated for infusion of either saline or the NE inhibitor ONO-5046 (50 mg kg–1 bolus followed by continuous infusion of 50 mg kg–1 hour–1 using a syringe pump), the cremaster muscle was exteriorized and leukocyte responses quantified by intravital microscopy. (A,B) Leukocyte responses of firm adhesion and transmigration at 4 hours after IL-1ß administration in the different groups of animals. The data shown are the means±s.e.m. from samples of 4-12 mice per group. Statistically significant differences in responses between mice receiving i.s. saline and mice receiving i.s. IL-1ß are shown by asterisks: **P<0.01; ***P<0.001. In mice receiving i.s. IL-1ß, differences to animals injected with i.v. IgG-saline (control group) are shown by crosses:+P<0.05; +++P<0.001. (C-E) IL-1ß-stimulated cremaster muscles from mice injected with either i.v. IgG-saline (C) or GoH3/ONO-5046 (D,E) were immunostained with specific mAbs for expression of PECAM-1 (APC; green), laminin 10 (Alexa Fluor 555; red) and CD11b (Alexa Fluor 488; white). Samples were observed at RT using a Zeiss LSM 5 Pascal confocal laser-scanning microscope (using a 40x water-dipping achroplan objective with a numerical aperture of 0.75) equipped with argon (excitation wavelength of 488 nm) and helium-neon (excitation wavelengths of 543 nm and 633 nm) lasers; the figure shows merged images captured from the three channels used. (E) A high-magnification image of the specified vessel segment shown in D in order to illustrate better the localization of leukocytes within the vessel wall. This is further indicated with an accompanying schematic diagram in which the position of endothelial cells (as identified using an anti-PECAM-1 mAb) is shown by a green line, the position of the basement membrane (as identified by an anti-laminin-10 Ab) is shown by a red line and trapped leukocytes (identified using an anti-CD11b mAb) are shown in white. All images are from tissue samples stained and analysed blind with at least nine random fields per tissue being captured from three or four mice in each group. Bars, 10 µm.