fMLP-stimulated Lsc KO neutrophils are less adherent. (A) Neutrophils plated on either plastic or fibronectin were stimulated with 10 μM fMLP for 30 minutes and washed 3 times, and the remaining adherent neutrophils were quantified as a fraction of the total plated. Data are the mean ± SEM calculated from 6 independent samples in each experiment. Plots are representative of at least 2 independent experiments. (B) The adherent area of WT and Lsc KO neutrophils plated on a glass coverslip and stimulated with 10 μM fMLP in a Zigmond chamber are as described in Figure 3A. Lsc KO neutrophils are less spread out than WT neutrophils. (C) Surface expression of CD18 (β₂-integrin) and CD29 (β₁-integrin) on fMLP-stimulated WT and Lsc KO neutrophils. Neutrophils in suspension were incubated with 10 μM fMLP or vehicle alone for 30 minutes and labeled with an anti-CD18 or anti-CD29 antibody, and the geometric mean cellular fluorescence (MCF) of the labeled neutrophils was measured by flow cytometry and normalized to the MCF of unstimulated WT neutrophils. Data are the mean normalized MCF ± SEM from 3 independent experiments. CD18 expression is similar in WT and Lsc KO neutrophils before and after stimulation. CD29 expression is reduced in Lsc KO neutrophils before and after fMLP-stimulation.