TABLE 1.
Sequences of oligonucleotides used in this study
The oligonucleotide sequences used for PCR, site-directed mutagenesis, methylation status of hIDE core promoter, and gel retardation experiments (28) are given.
| Method | Primer | Sequence (5′–3′) (restriction site) |
|---|---|---|
| PCRa | OUTER | GCAATATTTGGAGGATCCGACAAT |
| R147 | TCTCTTGATGGCTGGATTATTCAT | |
| R59 | ACTGAGCGGAAGGTGCTGGGCAGT | |
| R98 | TTGGAAAACCCATTCTTGAGG | |
| F (−1209) | GTATGGTACCCTCTGCATTGGGACTG (KpnI) | |
| F (−575) | GTATGGTACCCACGCCGCACTCGA (KpnI) | |
| F (−118) | GTATGGTACCTCTCCGCAGCTCC (KpnI) | |
| F (−82) | GTATGGTACCGCTAGAGCATGCGC (KpnI) | |
| F (−19) | GTATCTCGAGCCGGCGACTGCGCTGG (XhoI) | |
| R (−19) | GACTAAGCTTGATCACCGCAAACGCT (HindIII) | |
| R (−59) | GACTAAGCTTCTGCGCACTGCGCA (HindIII) | |
| Site-directed mutagenesisb | NRF-1 mut1 | GCCGCGGCTAGAAAATGCGCAGTGCGCAGGGCCGGCTCGAAGCGC |
| NRF-1 mut2 | GCCGCGGCTAGAGCATGAACAGTGCGCAGGGCCGGCTCGAAGCGC | |
| Methylation status of hIDE core promoter | CpG-F | GGAAAGTCTGGGTGCTGGCTC |
| CpG-R | TTAAGTGCTGAATCACCACTTTGCAAC | |
| EMSA | IDE WT | CCGCGGCTAGAGCATGCGCAGTGCGCAGGG |
| IDE MUT | CCGCGGCTAGAAAATGAACAGTGCGCAGGG |
a Restriction site sequences are shown in italics and underlined.
b Mutated bases are shown in bold and underlined.