Figure 7. Melatonin enhanced fisetin-mediated inhibition of NF-κB binding and acetylation.

(A, B), Human MeWo cells were treated with 20 µM fisetin (F) and 1.0 mM melatonin (MT). The binding of p50 and p65 to th biotinylated COX-2 promoter probe (A) was ananlyzed by pulldown and Western blot, the relative binding activities of p50 and p65 to COX-2 promoter (B) was analyzed by densitometric analysis. (C, D), MeWo cells were treated with F (20 µM) and MT (1.0 mM). Nuclear lysate from the treated cells was immunoprecipitated with antibodies against p50 and p65. The acetylated levels of p50 and p65 (C) was analyzed by Western blot, and the acetylated levels of p50 and p65 were quantitatively calculated by densitometric analysis (D).