TABLE 1.
Oligonucleotide sequences for PCR and EMSA used in this study
F indicates forward, and R indicates reverse.
| Target gene (product size) | Primer (5′ → 3′) |
|---|---|
| Cloning for rat SelW promoter | |
| −973SelW/Luc, F | GCGCGGTACCGCCTTGCGCTTCCTAGGC |
| −240SelW/Luc, F | GCGCGGTACCGAAGGGACAGCGAGGGGC |
| +32, R | CCGCAAGCTTGCACAAAGCGAGGACCCG |
| Site-directed mutagenesis for rat SelW promoter | |
| E1m, F | GATTTTTTGTTTTGAGACGACAGCTCTTGTAGCCCAG |
| E1m, R | CTGGGCTACAAGAGCTGTCGTCTCAAAACAAAAAATC |
| E2m, F | CTTCAGTCTCCTCTTCCCGAAAGCTGGGATTAGAGGC |
| E2m, R | GCCTCTAATCCCAGCTTTCGGGAAGAGGAGACTGAAG |
| E3m, F | GGCAAGTCCTCTGCGCCGACAGCGCTACAGCCCAAGC |
| E3m, R | GCTTGGGCTGTAGCGCTGTCGGCGCAGAGGACTTGCC |
| E4m, F | GGAGGAGAGATCCATCCCGATAGTCTCCCGATGCGTG |
| E4m, R | CACGCATCGGGAGACTATCGGGATGGATCTCTCCTCC |
| Gene amplification | |
| SelW, F (232 bp) | GTGTATTGTGGAGCTTGAGGC |
| SelW, R | CCAAGGCAGCTTTGATGGCGG |
| MyoD, F (482 bp) | CATCCGCTACATCGAAGGTC |
| MyoD, R | TCGCATTGGGGTTTGAGCC |
| p21, F (295 bp) | GTCCAATCCTGGTGATGTCC |
| p21, R | CAGGGCAGAGGAAGTACTGG |
| Myogenin, F (430 bp) | AGGAGAGAAAGATGGAGTCCAGAG |
| Myogenin, R | TAACAAAAGAAGTCACCCCAAGAG |
| MyHC, F (625 bp) | AGAAGGAGGAGGCAACTTCTG |
| MyHC, R | ACATACTCATTGCCGACCTTG |
| TrxR1, F (876 bp) | TGGATTTTGTCACACCGACTCC |
| TrxR1, R | CGATGGCGTAGATGTAAGGCAC |
| Hprt1, F (225 bp) | GCAAACTTTGCTTTCCCTGG |
| Hprt1, R | GCTTTGTATTTGGCTTTTCC |
| Gapdh, F (369 bp) | CATGACAACTTTGGCATTGTG |
| Gapdh, R | GTTGAAGTCGCAGGAGACAAC |
| EMSA | |
| E1 | GTTTTGAGACAACTGCTCTTG |
| E2 | CTCTTCCCAAATGCTGGGAT |
| E3 | CTCTGCGCCAACTGCGCTACAGCC |
| E4 | GATCCATCCCAATTGTCTCCCGAT |
| MCK | CCCCCCAACACCTGCTGCCTGA |
| ChIP | |
| E1, F | CCTCGAACCCACAGAGATTTC |
| E1, R | GAAGAGGAGACTGAAGGATTG |
| Gapdh, F | AGCTACTCGCGGCTTTACG |
| Gapdh, R | TCACCTGGCACTGCACAAG |