E‐selectin and ICAM‐1 are incorporated into detergent‐insoluble membrane domains following clustering in endothelial cells

Here we present data supporting the role of lipid rafts in endothelial cells during leukocyte adhesion. Following adhesion of THP-1 cells or antibody-mediated clustering, both E-selectin and intercellular adhesion molecule-1 (ICAM-1) partitioned into the detergent-insoluble portion of the endothelial cellular lysate. Sucrose gradient centrifugation revealed the partitioning of clustered E-selectin and ICAM-1 with the low-density fraction where they co-fractionated with src family kinases, markers of lipid rafts. Depleting the plasma membrane of cholesterol inhibited clustering of adhesion molecules following their antibody-induced crosslinking and inhibited their association with src kinases. Thus, our data suggest that E-selectin and ICAM-1 associate with lipid rafts in human endothelial cells following leukocyte adhesion.