Figure 3

Genistein increases p27kip1 expression and promotes FOXO3 binding to the p27kip1 promoter. (A) Protein from control and genistein treated HT-29 cells was separated and immunoblotted for p27kip1. Immunoblots revealed that genistein increased p27kip1. The experiment was performed three independent times and quantified using densitometry (n = 3, *p < 0.05). (B) Genistein inhibits FOXO3 disassociation from p27kip1 promoter. ChIP assay was performed to determine FOXO3 binding to p27kip1 promoter (-110 bp). EGF-induced FOXO3 disassociation from p27kip1 promoter (-110 bp) was inhibited by genistein. Input represents PCR amplification of DNA from cell lysate prior to immunoprecipitation with the primers from p27kip1 promoter. This experiment was repeated two times.