Effect of TNF-α on IκB expression and NF-κB DNA binding.

<p>In (A) is shown the effect of TNF-α (100 ng/ml) on the expression of IκBα in isolated human eosinophils. Incubations were terminated at the indicated time-points after addition of TNF-α (top panel) or medium in the simultaneously prepared control cells isolated from the same donor (lower panel). The data are representative of three separate experiments using eosinophils isolated from different donors. In (B) is shown the effect of TNF-α on NF-κB DNA binding in isolated human eosinophils. Incubations were terminated at the indicated time-points after addition of TNF-α (100 ng/ml). NF-κB DNA-binding activity was analyzed by electrophoretic mobility shift assay. Arrowheads indicate the different specific bands. In (C) is shown the total optical density (OD; arbitrary units) values of the abovementioned three specific bands (mean ± SEM) from five experiments using eosinophils isolated from different donors. * indicates P<0.05 as compared with the respective value at 0 min timepoint.