Lipid raft disruption by mβCD inhibits neutrophil rolling on P-selectin, E-selectin, or PSGL-1 but does not prevent firm adhesion of fMLP-activated neutrophils on ICAM-1. (A) Freshly isolated neutrophils, treated for 10 minutes with 10 mM mβCD, were perfused on P-selectin (▪), E-selectin (▦), or PSGL-1/μ (□). Results of 3 representative experiments are shown (mean ± SEM). ***P < .001. (B) Neutrophils treated with proteinase K (1.7 μg/mL for 20 minutes at 37°C; ▦) or its vehicle (▪) were perfused on P-selectin, E-selectin, or PSGL-1/μ chimera. (C) Neutrophils were treated for 10 minutes with 10 mM mβCD (▪) or its vehicle (○). After resuspension (10⁶ cells/mL) in medium supplemented with fMLP (10 nM), neutrophils were perfused for 3 minutes under shear stress (0.5 dyn/cm²) on a coverslip coated with ICAM-1. After a pause of 3 minutes, cell detachment was induced by perfusion of cell-free medium at various shear stress levels. Results (mean ± SEM; n = 3) are shown as percent of the number of adherent cells remaining after perfusion of cell-free medium at 0.5 dyn/cm².