Effect of lipid raft disruption by mβCD on cell surface expression of PSGL-1, L-selectin, or sLex. (A) Neutrophils were treated with mβCD (10 mM, 10 minutes; plain histograms) or its vehicle (bold lines), stained with antibodies against PSGL-1, L-selectin, or sLex using PE-labeled goat antimouse as secondary antibody, and analyzed by flow cytometry. Histograms are representative of 9 independent experiments. (B) KG1 cells were treated with mβCD (10 mM) or its vehicle and then immediately processed for electron microscopy. Left panels: the scale bar represents 0.7 μm (×14 000). Middle panels: the scale bar represents 0.2 μm(×50 000). Arrowheads indicate immunogold labeling of PSGL-1. Isotypic control is shown in the right panel: the scale bar represents 0.3 μm (×30 000). (C) Distribution of PSGL-1 (panel B, middle panels): percentages of gold particles in cell body (□) or microvilli (▪). Mean values plus or minus SEM of 150 counts is shown (24 microscopic fields).
