Figure 5

Methylation of CpG-4 decreases USF1/2-associated complex stability. EMSAs were performed with 32P-end-labeled probes. Nuclear protein extracts (5 μg) from HT29 cells were pre-incubated in the presence of either no competitor or 10-, 50-, 100-fold molar excess of cold competitors (unmethylated and methylated) or oligonucleotides including either a consensus binding site for HNF1-alpha or USF1/2. In methylated probes, dC nucleotide in CpG dinucleotides is substituted by an internal methyl deoxyCytidine.