Chemerin Does Not Inhibit Inflammatory Responses Figure 1. Effect of chemerin on the production of cytokines by activated mouse macrophages. Peritoneal macrophages from WT mice, collected after i.p. thioglycolate injection, were exposed or not for 1 h to 100 nM recombinant mouse chemerin. The cells were further stimulated by 100 ng/ml LPS. Unstimulated cells were used as controls. (A) At the indicated time points, cells were harvested for the determination of transcript levels for IL-6, TNF-a and IL-10, using quantitative RT-PCR. Results are expressed as mean 6 SEM for 2 independent experiments performed in duplicate. (B) At the indicated time points, IL-6, TNF-a and IL-10 levels were determined in the supernatants by ELISA. Results are expressed as mean 6 SEM for 3 independent experiments performed in duplicate. doi:10.1371/journal.pone.0040043.g001 luminescence units) were normalized to the response to 20 mM ATP, and the parameters of the dose-response curves were determined with the PRISM software using nonlinear regression applied to a sigmoidal dose-response model. Chemotaxis assay Chemotaxis assays were performed on peritoneal macrophages (Bio-Gel procedure) as previously described [7]. Briefly, migration PLoS ONE | www.plosone.org 3 June 2012 | Volume 7 | Issue 6 | e40043