Figure 2.

Cdc42 regulates neutrophil activation and secretory vesicle degranulation. Freshly isolated human neutrophils were treated with DMSO (solvent control, white bars) or 10 μM casin (black bars) for 30 min at 37°C. Cells were left untreated or stimulated with 100 ng/ml LPS, 10 nM C5a, or 1 μM fMLP. Neutrophils were stained with antibodies to the activation marker CD62L (FITC), and the degranulation markers CD11b (PE), and CD18 (APC), respectively. Displayed are representative flow cytometry histograms of unstimulated PMNs (red) or cells stimulated with 100 ng/ml LPS (blue). (A) Neutrophils were gated according to the size and granularity shown in the sideward- (SSC) and forward-scatter (FSC). (B–D) Neutrophils were gated for the activation marker CD62L (B) and the degranulation markers CD11b (C) and CD18 (D). CD11b^high, CD62L^high, and CD18^high cells were used for analysis. Bar graphs represent the ratio (% ± SD) from three independent experiments of CD62L^high (E), CD11b^high (F), and CD18^high (G) cells assessed by flow cytometry. ***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; unpaired, two-tailed students t-test.