Figure 5

Effect of galangin on the TPA-induced the DNA-binding activity of NF-κ B and AP-1/expressions of NF-κ B, c-Fos, c-Jun/Iκ Bα phosphorylation and degradation in HepG2 cells. Nuclear extracts were prepared from HepG2 cells that treated with various concentrations of galangin (0, 1, 2.5, and 5 μM) in the presence of TPA (70 nM) for 12 h, and then used to analyze (A) NF-κB and (B) AP-1 DNA-binding activity by EMSA, as described in “Materials and methods” section. Lane 1: nuclear extracts incubated with 100-fold excess unlabeled consensus oligonucleotide (comp.) to confirm the binding specificity. Lane 2 represents nuclear extract from HepG2 cells in the absence of TPA (negative control). (C) Nuclear or cytosolic extracts were subjected to SDS-PAGE followed by western blotting with specific antobodies (anti-NF-κB, anti-c-Fos, anti-c-Jun anti-p-IκBα, anti-IκBα). C23 and β-actin were used as internal control.