Figure 7.

Effects of SKF96365 and 2-APB on the direct polarization of neutrophils and CSE gradient in the Zigmond chamber. (A) Neutrophils were given 30-minutes pretreatment with phosphate buffered solution, 40 µM of SKF96365, or 100 µM of 2-APB, and then assembled in Zigmond assays. Time-lapse microscopy was used to record cell morphology on the bridge at a 30 s interval and the direction of the fMLP gradient (0–100 nmol/L) was indicated by the wedge, and cell images were acquired at 20× object by optical microscope. The icon above the images indicates the direction of fMLP gradient. A higher magnification image of the boxed section is shown below each panel of images for a clearer view of cell polarization. The percentage of cells polarized in each panel was quantified as described under the “Methods” section. Images were acquired at 20× objective. (B) The data are expressed as the mean ± SD from 3 independent experiments. CSE, cigarette smoke extract; fMLP, N-formyl-methionine-leucine-phenylalanine.