Figure 4

Genistein increases FOXO3-p53(mut) interaction in HT-29 cells. (A) Protein from control and genistein treated HT-29 cells was immunoblotted for p53. Genistein increased p53 expression 2.5-fold. The experiment was performed three independent times and quantified using densitometry (n = 3, *p < 0.05). (B, C) Protein from different experimental groups from HT-29 cells was immunoprecipitated with antibody against FOXO3 (mouse) and immunoblotted using antibody against p53 (rabbit). Co-immunoprecipitation revealed that genistein increased FOXO3-p53(mut) interaction and prevented EGF induced disassociation of p53 and FOXO3. IgG (mouse) was used as a negative control. Experiments were repeated three times. (D) Protein from HCT116 with wild type p53 was immunoprecipitated with anti-FOXO3 (mouse) and immunoblotted using antibody against p53 (rabbit). Nether EGF nor genistein affected FOXO3 interaction with wild type p53 in HCT116 cells.