FIG. 4.

SFN inhibited the activation of the nuclear factor-κB (NF-κB) pathway induced by TNF-α stimulation. (A) ECV 304 cells were pretreated for 18 h with indicated concentrations of SFN and stimulated with TNF-α subsequently. Nuclear extracts from each group were subjected to the electrophoretic mobility shift assay (EMSA) for analyzing the DNA binding activity of NF-κB. (B) The cells were preincubated with SFN and stimulated with TNF-α for the indicated periods. Cell lysates from each group were examined by immunoblotting analysis using p-IκBα and IκBα antibodies. (C) Cell lysates from each group were examined by immunoblotting analysis using p-IKKβ and IκB kinase (IKK)β antibodies. At least three independent experiments were performed, with similar results.