Figure 5
Elimination of 12/15-LO leads to a reduced response of neutrophils to CXCL1. (A) Flow cytometry analysis of CXCR2 expression was performed on peripheral blood neutrophils from WT and Alox15-/- mice under baseline conditions. Whole blood was stained for CD45, 7/4, GR-1 and CXCR2. CXCR2 expression on the surface of peripheral blood neutrophils was determined by using flow cytometry. (n = 3). (B) Flow cytometry analysis of CXCR2 expression on isolated neutrophils treated with or without CXCL1 (10 pg/ml) for one hour (n = 6). Migration of labeled neutrophils from WT mice (circles) and Alox15-/- mice (triangles) toward different concentrations of CXCL1 (C) or fMLP (D) was investigated in an in vitro chemotaxis assay. Neutrophils were applied on the top of the filter and allowed to migrate toward the chemoattractant for one hour at 37°C. Migrated cells were measured by fluorescence intensity. Chemotactic index was calculated as the ratio of migrated cells with chemoattractant to migrated cells without chemoattractant and ED50 was determined (n = 3).