Decreased neutrophil infiltration and MIP-2 production of CIA mice treated with anti-Cyr61 antibody. (A) Immunohistochemistry of joint tissue from CIA mice treated with 093G9 or con-IgG1. Left panel: Representative photomicrographs showing H & E staining (Up, 100×) and infiltrating neutrophils stained with mouse neutrophil marker anti-Gr1 Ab (Lower, 400×) in joint tissue from CIA mice treated with con-IgG1 or 093G9. Arrow points to representative neutrophils (red). Right panel: The number of infiltrating neutrophils in joint tissue from CIA mice treated with con-IgG1 (open bar, n = 6) or 093G9 (black bar, n = 6) were counted in six randomly selected fields per tissue slice by optical microscope. (B) MIP-2 mRNA relative expression in synovial tissues from CIA mice treated with control IgG1 (open bar, n = 6) or 093G9 (black bar, n = 6) was evaluated by real-time PCR. (C) Left panel: MIP-2 protein level in synovial tissues from CIA mice treated with control IgG1 (n = 4) or 093G9 (n = 4) was detected by western blotting. Right panel: The density of each band was quantified by densitometric scanner and the relative amount of MIP-2 protein was calculated from the ratio of each band value to β-actin. Data represent the mean ± SEM from three independent experiments. * P <0.05, ** P <0.01. CIA, collagen-induced arthritis; SEM, standard error of the mean.