Adhesion in the flow chamber and transmigration of ST3Gal-IV−/− neutrophils in vitro. The number of rolling and adherent leukocytes from ST3Gal-IV−/− and WT mice was determined in blood-perfused microflow chambers. (A) Rolling and (B) adherent leukocytes in autoperfused flow chambers (5.94 dynes/cm2) coated with 20 μg/ml P-selectin and 15 μg/ml ICAM-1 with or without 15 μg/ml CXCL1. Data in A and B are presented as mean ± SEM from at least four mice and four flow chambers. *, P < 0.05 versus WT mice (B). (C) The percentage of transmigrated neutrophils from WT control mice, ST3Gal-IV−/− mice, and WT control mice pretreated with neuraminidase was assessed in a transwell assay in which transwells were coated with an immortalized endothelial cell line, b.End5. In addition, transmigration of WT control neutrophils through a neuraminidase-pretreated b.End5 monolayer was analyzed. Data in C were determined from at least three independent experiments per group. *, P < 0.05 versus untreated WT control neutrophils (C). FOV, field of view.
