Binding of soluble PSGL-1, E-selectin/μ, or P-selectin/μ to neutrophils is inhibited by lipid raft disorganization. Neutrophils were treated with mβCD (10 mM, 10 minutes; plain histograms) or its vehicle (bold lines) and stained with PSGL1/μ, P-selectin/μ, or E-selectin/μ (5 μg/mL) preincubated with FITC-conjugated rabbit anti–human IgM. Binding was assessed by flow cytometry. A total of 5000 cells was analyzed in each experiment. Ligand binding was abolished by 5 mM EDTA (thin lines) specifically by anti–PSGL-1 mAb KPL1, anti–P-selectin mAb WAPS12.2, or anti–E-selectin mAb H18/7 (not shown). Histograms are representative of 9 to 14 experiments. Mean values of MFI and percentage of positive cells (n = 9-14) are indicated below the histograms.