FIG. 6.
Mutation of a potential Tcf/LEF binding site abolishes shifted DNA-protein complexes in an EMSA. Duplex oligonucleotides containing wild-type (WT) or mutated T2 sites (from 5"-CTTTGAT-3" to 5"-CTTTCGC-3") were incubated with 293T cell nuclear extract (lanes 2 to 9), and DNA-protein complexes were separated in 5% native polyacrylamide gels. No shifted band was detected without nuclear extract (lanes 1 and 10). Mutation of site T2 caused a clear reduction in the shifted bands (compare lanes 2 and 9). An increasing amount of wild-type cold oligonucleotide efficiently eliminates the shifted bands while mutated oligonucleotide does not (compare lanes 3 to 5 with lanes 6 to 8).