Hs2st^f/fTie2Cre⁺ mice show reduced neutrophil rolling velocity. (A) Rolling velocity of neutrophils in cremaster muscle venules was assessed by intravital microscopy. Up to 10 individual leucocytes were manually tracked over time using MATLAB software, and the velocity of the leucocytes was calculated. Each dot represents the cumulative frequency (summation of the velocity frequency and all frequencies below it). (B) Quantification of the rolling velocity of neutrophils in Hs2st^f/fTie2Cre⁺ mutant mice and wild-type littermate control (n = 6-10). (C) Neutrophil rolling was assessed in vitro using flow chambers. Endothelial cells were grown to confluence on glass slides, and bone marrow-derived wild-type neutrophils were infused through the slides under a shear stress of 1 dyne/cm². The rolling velocity of neutrophils was quantified with or without pretreatment with TNF-α (n = 3). (D) The number of rolling neutrophils was quantified. In a separate experiment, endothelial cells were pretreated with TNF-α (n = 3 for each experimental condition). Error bar represents SEM.