Figure 3.

Electrophoretic mobility shift assays (EMSA) of the oligonucleotide containing a -1565C>T change with human liver nuclear extract (NE). EMSAs demonstrate the binding of human liver nuclear proteins (indicated by arrows) to wild-type oligonucleotide probe containing -1565C only. EMSAs were performed as described in Methods. Oligonucleotides for wild-type (WT) and MT-1 (-1565T) were labelled with [³²P]-dATP. Oligonucleotides were incubated with nuclear extract at 4°C for 30 min, followed by 5% polyacrylamide gel electrophoresis. Nonradiolabeled cold competitors (CC) were added at 20 and 50-fold excess to determine the specificity of binding complex.