Table 4.

The sequences of biotinylated oligonucleotides used for DNA/protein binding assays

Binding Site	Lef-1 promoter position (bp)a	5′ Biotinylated Synthetic Oligonucleotide Sequenceb
T1	−2693/−2687	Biotin-5′-GGGGATCctttgtaAAGTTTTAAAAATTGC-3′
S1	−2670/−2640	Biotin-5′-TTCCTAGTAAattcttattCCCAAGAAGTA-3′
T1/S1	−2693/−2645	Biotin-5′-GGGGATCctttgtaAAGTTTTAAAAATTGCTTCCTAGTAAattcttattCCCAAG-3′
T2	−2427/−2421	Biotin-5′-CATTTCTTTATGTCctttgttTACTGTTCTG-3′
T3	−2254/−2248	Biotin-5′-CTCCTGTctttgtaCAAAATTCATC-3′
T4	−2240/−2234	Biotin-5′-CAAAATTCatcaaagAGACATGG-3′
T3/T4	−2254/−2234	Biotin-5′-CTCCTGTctttgtaCAAAATTCatcaaagAGACATGG-3′
S2	−2956/−1951	Biotin-5′-GAAAGATATattgttTAAAAAGC-3′
T5	−921/−915	Biotin-5′-CTCGGCCGGGaacaaagAGGGGTCGG-3′
S3	−780/−760	Biotin-5′-CAAAAActttattcttGGCA-3′
Mutated T1	−2693/−2687	Biotin-5′-GGGGATCagggtgcAAGTTTTAAAAATTGC-3′
Mutated S1	−2670/−2640	Biotin-5′-TTCCTAGTAAcggaggcggCCCAAGAAGTA-3′
Mutated T1/S1	−2693/−2645	Biotin-5′- GGGGATCagggtgcAAGTTTTAAAAATTGCTTCCTAGTAAcggaggcggCCCAAG-3′
Mutated T2	−2427/−2421	Biotin-5′-CATTTCTTTATGTCagggtggTACTGTTCTG-3′
Mutated T3/T4	−2254/−2234	Biotin-5′-CTCCTGTagggtgcCAAAATTCcgaccctAGACATGG-3′
Mutated S2	−2956/−1951	Biotin-5′-GAAAGATATcggtggAAAAAGC-3′
Mutated T5	−921/−915	Biotin-5′-CTCGGCCGGGccacccctAGGGGTCGG-3′
Mutated S3	−780/−760	Biotin-5′-CAAAAAagggcggaggGGCA-3′

^aBase pair position is relative to the translational ATG start site (+1) in exon 1 of the human Lef-1 gene.

^bSynthetic oligonucleotides containing the wild type or mutant sequences of putative TCF and/or Sox binding sites in the human Lef-1 promoter were labeled at the 5′ end with biotin. The potential binding sequences (wild type and mutated) are shown in lowercase and bold.