Impaired transendothelial migration of neutrophils in EHCgp130^−/−mice after challenge with TNF-α or thioglycollate. Neutrophil rolling flux fraction (A), rolling velocity (B), firm adhesion (C), and transendothelial migration (D) were measured in venules of cremaster muscle from WT and EHCgp130^−/− mice 2 hours after TNF-α was injected into the scrotum. (C) As indicated, PTx was injected intravenously before TNF-α was injected. (E) Three-dimensional reconstructions of confocal microscopic images of TNF-α–stimulated venules of cremaster muscle from WT and EHCgp130^−/− mice. Neutrophils (green) were stained with anti–neutrophil elastase antibody, and endothelial cells (red) were stained with anti-CD31 antibody. The bar represents 10 µm. (F) Electron micrographs of TNF-α–stimulated venules of cremaster muscle from WT and EHCgp130^−/− mice. For each genotype, the right panel is a higher magnification of the area within the dashed lines in the left panel. N, neutrophil; P, pericyte, EC, endothelial cell. The bar in the left panel represents 10 µm, the bar in the right panel represents 2 µm. (G-H) Mice of the indicated genotype were injected with thioglycollate intraperitoneally. After 4 hours, peritoneal cells were lavaged and neutrophils were quantified by flow cytometry. The data in (A-D) represent the mean ± SEM from 15 to 20 venules from 4 to 5 mice in each group. The data in (E-F) are representative of at least 10 randomly selected vessel segments. The data in (G-H) represent the mean ± SEM from 5 mice in each group. *P < .05, **P < .01.