Fig. 4

Higher number of pro-inflammatory cells in circulation and joints of M3R-deficient mice. a, b Quantification of FACS analysis of total CD45+ (a) and CD45 + Ly6C + GR1+ (b) cells in whole blood from male LPS control and arthritic M3R^−/− and WT animals. Data are given as box plots with the median indicated by a solid line within the box. *P < 0.05 CAIA vs. respective LPS control; ^# P < 0.05: M3R^−/−-CAIA vs. WT-CAIA; ^§ P < 0.05: M3R^−/−-LPS vs. WT-LPS. c–f Immunohistochemical staining for neutrophils in knee joints of male WT-LPS (c), M3R^−/−-LPS (d), WT-CAIA (e), M3R^−/−-CAIA (f) mice. Original magnification: 200×. g Quantification of neutrophil staining with data given as average number of positive cells per high-power field (hpf) ± SEM. *P < 0.05 CAIA vs. respective LPS control. h Real-time RT-PCR analysis of Cxcl2 mRNA expression in paws of male WT and M3R^−/− mice with CAIA and respective LPS-treated control mice, normalized to β-actin mRNA. Data are given as –dCT values presented as box plots. *P < 0.05 and **P < 0.01 CAIA vs. respective LPS control. i Correlation of Cxcl2 mRNA expression with the arthritis score of the respective paw in arthritic WT and M3R^−/− mice. Dotted lines indicate the mean Cxcl2 mRNA expression of the corresponding LPS-treated control mice. CAIA collagen antibody-induced arthritis, LPS lipopolysaccharide, M3R M3 muscarinic acetylcholine receptor, WT wild-type