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Blood, Vol. 107, Issue 5, 2101-2111, March 1, 2006
Dynamic shifts in LFA-1 affinity regulate neutrophil rolling, arrest, and transmigration on inflamed endothelium
Blood Green et al.
107: 2101
Supplemental materials for: Green et al
Files in this Data Supplement:
- Figure S1. Shape characteristics and adhesion receptor expression on isolated human neutrophils (JPG, 180 KB)
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A) Freshly isolated human neutrophils were examined for forward scatter and side scatter characteristics by FACScan flow cytometry at 23°C, after 10 minutes at 37°C and following 10 minutes with 1 µM fMLP at 37°C. Forward scatter vs side scatter dot plots are representative of the average population response from >50 separate experiments. B) The expression of Mac-1 and high affinity CD18, as detected by activation epitope reporter 327C, was measured by FACScan following the protocols described in A). Neutrophil suspensions at 23°C, 37°C and 37°C + fMLP were fixed and labeled with fluorescent antibody for 20 min prior to flow cytometry. Fluorescence histograms are representative of the average population response from >50 separate experiments. C) Isolated neutrophils were injected into a static adhesion chamber over unstimulated (left) and IL-1 activated (right) HUVEC monolayers. Images are 20× phase contrast after 5 minutes of interaction. Neutrophils above the monolayer appear phase bright while transmigrated neutrophils appear phase dark. Images are representative of the average response from >10 separate experiments.
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