Figure 5

Effect of genistein (Gen) on TPA-induced NF-κB and AP-1 activation and translocation of p65 in hepatoma cell lines. After 45 min, EMSAs of nuclear extracts of cells were performed. TPA induced AP-1 (A, B, C) and NF-κB (D, E, F) activation, which was inhibited by Gen and different inhibitors (B, E). For translocation of p65, cells were pretreated with 20 μM Gen with or without inhibitors for 45 min before incubation with TPA for 45 min. Then, nuclear and cytosolic extracts were prepared and subjected to western blotting with antibodies specific for p65 translocation (G). Cells were pretreated with 20 μM Gen with or without inhibitors for 30 min before incubation with TPA for 45 min, and then nuclear extracts were prepared as described in the Methods section.