Click on image to view larger version.
Figure 1. Transwell migration of peripheral neutrophils. (A) The data are the mean of a representative experiment conducted in triplicate (± SD) with cells isolated from different healthy adult volunteers. Il-8 was introduced in the lower chamber. S100A8 was introduced in the lower or upper chamber of the Transwells. Analysis of the data indicated a dose-dependent increase in neutrophil migration to the lower chambers of the Transwells when IL-8 and S100A8 were added to the lower and upper chambers, respectively. The addition of S100A8 to the lower wells did not have any measurable effect at the concentrations and conditions tested (#p < 0.05; *p < 0.01 when compared with control with no compound added). (B) The data represent the average of 4 experiments performed in duplicate (± SD) with cells isolated from different adult volunteers. S100A8 was introduced in the lower, upper, or both chambers of the Transwells at a concentration of 10–9 M. Analysis of the data indicated that the increased migration of neutrophils observed when S100A8 was added to the upper well was inhibited by the addition of S100A8 at an equimolar concentration in the lower well (*p < 0.01 when compared with control with no compound added). (C) The data represent the average of 2 experiments performed in duplicate (± SD) with cells isolated from different adult volunteers. S100A8 was added to the upper chamber of the Transwell at a concentration of 10–9 M. Il-8-mediated chemotaxis served as a positive control for the effect of pertussis toxin and was added to the lower chamber of the Transwell at a concentration of 10–9 M. The cells were incubated with different concentrations of pertussis toxin for 30 min at room temperature before the beginning of the assay. Analysis of the data indicated that the fugetactic effect of S100A8 was pertussis-toxin-sensitive. Il-8-mediated chemotaxis displayed a similar sensitivity (*p < 0.01 when compared with the migration ratio with no pertussis toxin for IL-8 and S100A8 separately).
