Fig. 4.

Binding of WT1-pTj to p53, displacement of p53 bound to WT1, and WT1-pTj modulation of transcriptional activity of p53. (A) The interaction between p53 and WT1-pTj was determined using biotinylated peptide and CL-ELISA; ^*p < 0.01. (B) Competitive inhibition of WT1 association to p53 was evaluated in wells coated with recombinant p53, incubated with WT1 and then exposed to WT1-pTj or C PEP at indicated concentrations. Binding of p53 to WT1 was quantified with anti-WT1 monoclonal antibody; ^*p < 0.02 compared to C PEP effect. (C) A2058 cells were transfected with the p53-responsive luciferase reporter construct, and then treated with 0.5 mM WT1-pTj or 0.5 mM C PEP for 24 h. The ratio of firefly to Renilla luciferase was determined and normalized to the value obtained for untreated cells (=100%). Data are representative of two independent experiments. * p < 0.02 compared to Control (untreated) and C PEP systems.