Ab ligation of Ly6G does not affect markers of neutrophil activation or adhesion to inflamed endothelium. (A) Flow cytometric assessment of activation markers CD11b and CD62L on CD45⁺Gr-1⁺CD11b^hi cells (10 minutes). (B) F-actin as assessed by phalloidin staining after preincubation with anti-Ly6G, isotype, or PMA (10 minutes). (C) Kinetic staining for ROS. Panels A-C reflect the findings of at least 2 experiments. (D) Adhesion of purified BM neutrophils preincubated with anti-Ly6G or isotype to TNFα-activated bEND5 endothelial cells under low physiologic flow conditions (0.5 dyne/cm²). Shown is abrogation by EDTA consistent with calcium dependence of the neutrophil-endothelium interaction. Data are representative of 3 independent experiments.