Fig. 7.

Intervention of the MAP-kinase and NF-κB pathways. Neutrophils were activated with CSM and drugs as described in material and methods “Materials and methods.” Western blot analysis for MAPK pathway; Erk1/2, phospho p38 from whole cell extracts (50 μg) and c-fos (25 μg) from nuclear extracts (a) and NF-κB pathway; IκB-α from whole cell extracts and p65 from nuclear extracts (b) were carried out with related antibodies. Representative results of three independent experiments are shown. β-actin and Lamin A served as loading controls from cytoplasm and nuclear fractions, respectively. c Neutrophils were preincubated with S (10^–7 M) or FP (10^–9 M) or in combination for 90 min and then activated with CSM (0.06 OD) for 30 min and then the nuclear proteins were analyzed in triplicate for the DNA binding activity of NF-κB using the kit. NF-κB activation is evaluated with reading optical density at 450 nm. Values (mean±SEM) are representative data from one of five independent sets of experiments. Asterisk indicates significant differences between medium-treated cells and cells treated with CSM (*p < 0.05), and the number sign represents the significance differences between cells treated with CSM and S or FP (^# p < 0.05, ^## p < 0.01). S and FP indicate for salmeterol and FP, respectively