Oligonucleotides used for EMSA experiments based on the presence of predicted Wt1 binding sites in the minimal human EPO promoter
Oligonucleotide . | Position of mutations within the oligonucleotides . | Single-strand sequence . |
|---|---|---|
| Oligo -116/-87 | — | 5′-TGCTCTGACCCCGGGTGGCCCCTACCCCTG-3′ |
| Oligo -86/-67 | — | 5′-GCGACCCCTCACGCACACAG-3′ |
| Oligo -66/37 | — | 5′-CCTCTCCCCCACCCCCACCCGCGCACGCAC-3′ |
| Oligo -66/-37 | ||
| Mutant A | -60 to -58 | 5′-CCTCTCAAACACCCCCACCCGCGCACGCAC-3′ |
| Mutant B | -54 to -52 | 5′-CCTCTCCCCCACTTTCACCCGCGCACGCAC-3′ |
| Mutant C | -60 to -58 and -54 to -52 | 5′-CCTCTCAAACACTTTCACCCGCGCACGCAC-3′ |
Oligonucleotide . | Position of mutations within the oligonucleotides . | Single-strand sequence . |
|---|---|---|
| Oligo -116/-87 | — | 5′-TGCTCTGACCCCGGGTGGCCCCTACCCCTG-3′ |
| Oligo -86/-67 | — | 5′-GCGACCCCTCACGCACACAG-3′ |
| Oligo -66/37 | — | 5′-CCTCTCCCCCACCCCCACCCGCGCACGCAC-3′ |
| Oligo -66/-37 | ||
| Mutant A | -60 to -58 | 5′-CCTCTCAAACACCCCCACCCGCGCACGCAC-3′ |
| Mutant B | -54 to -52 | 5′-CCTCTCCCCCACTTTCACCCGCGCACGCAC-3′ |
| Mutant C | -60 to -58 and -54 to -52 | 5′-CCTCTCAAACACTTTCACCCGCGCACGCAC-3′ |
Oligonucleotide numbers indicate the positions of the nucleotides relative to the transcription start site. Transcription start site is given here at nucleotide 387 of the EPO gene (NCBI accession number M11319.1), whereas Blanchard et al12 describe the transcription start site at nucleotide 386.59 The underlined letters in the single-stranded sequences indicate the nucleotide mutations.