Primers used in this study for promoter cloning, mutagenesis, real-time PCR, EMSAs, and ChIP assays
PCR primers are numbered relative to the adenine base of the initiator codon (ATG), which is assigned as +1. Incorporated restriction sites of both MulI (ACGCGT) and KpnI (GGTACC) are underlined. Oligonucleotides are the same for both mutagenesis and EMSAs. Mutated sequences are highlighted in bold.
Primer | Nucleotide Sequence (5′ to 3′) |
|---|---|
| PCR and cloning | |
| 2B15-54For | AGCCATGGTACCGAGAAATGACAGAAAG |
| 2B15-101For | AGCCATGGTACCTAGCAGTTATATTTTAAC |
| 2B15-154For | AGCCATGGTACCTCCTTGGCATGCACCTATTC |
| 2B15-202For | AGCCATGGTACCCACCAAACACACTAAAA |
| 2B15-253For | AGCCATGGTACCCTAGGTCATAAAAATTA |
| 2B15-310For | AGCCATGGTACCGCCACTGTTCTTAACAGT |
| 2B15-361For | AGCCATGGTACCTACTTCTTCTCTCTATGTC |
| 2B15-412For | AGCCATGGTACCAAGACATTACTTGCACCACGA |
| 2B15-458For | AGCCATGGTACCTTGGTGAGTCATATGGC |
| 2B15-556For | AGCCATGGTACCGAGGCTATTGTATAGAATC |
| 2B15-595For | AGCCATGGTACCTAGGGTAGGATGGATG |
| 2B15-705For | AGCCATGGTACCAATCTAGTCCTGCATCC |
| 2B15-747For | AGCCATGGTACCTTCACAATGCAAGCCTCTG |
| 2B15-2716For | AAATCTGGGTACCACATAATGCCTGCACT |
| 2B15-3Rev | AGAACGCGTGGTCTTATGCAATGCTTCTT |
| Site-directed mutagenesis/EMSA | |
| 5′AP-1wt | GTACACACTAATTGGTGAGTCATATGGTATATG |
| 5′AP-1m | GTACACACTAATTGGCAGAGCATATGGTATATG |
| 3′AP-1wta* | GGTATATGAATTATGTGTCAACAAAGTTTTAGAAGAC |
| 3′AP-1ma* | GGTATATGAATTACAGAGCAACAAAGTTTTAGAAGAC |
| 3′AP-1wtb† | TATGAATTATGTGTCAACAAAGTTT |
| 3′AP-1mb† | TATGAATTACAGAGCAACAAAGTTT |
| 3′EREhwt | AAATAAATATGAGGTCATCAATCTTTTGTTG |
| 3′EREhm | AAATAAATATGAAAAAATCAATCTTTTGTTG |
| 5′EREhwt | CTTACATATTCTAGGTCATAAAAATTATTG |
| 5′EREhm | CTTACATATTCTATTTTTTAAAAATTATTG |
| EREfwt | CTCTCTATGTCAAGGGCACCGAACAGGC |
| EREfm | CTTCTTCTCTCTAAAAAGAGGGCACCGAAC |
| X. laevis VitellogeninA2 vitERE | AAAGTCAGGTCACAGTGACCTGATCAAAG |
| Quantitative real-time PCR | |
| c-Fos-For | TGATGACCTGGGCTTCCCAG |
| c-Fos-Rev | CAAAGGGCTCGGTCTTCAGC |
| c-Jun-For | CTTCAACCCAGGCGCGCTGAGCA |
| c-Jun-Rev | GTCTGAGGCTCCTCCTTCAGGGCCT |
| ERα-For | TTCGGCTCCAACGGCCTGGGGGGTTT |
| ERα-Rev | GGTACTGGCCAATCTTTCTCTGCCACCCT |
| Fra-2-For | TAGATATGCCTGGCTCAGGCAGTGCA |
| Fra-2-Rev | TCTTGATCACGCCAGGTCTTGGGA |
| 18S rRNA-For | CGATGCTCTTAGCTGAGTGT |
| 18S rRNA-Rev | GGTCCAAGAATTTCACCTCT |
| 1A3-For | ATGGCAATGTTGAACAATATG |
| 1A3-Rev | GGTCTGAATTGGTTGTTAGTAATC |
| 1A7-For | TGGCTCGTGCAGGGTGGACTG |
| 1A7-Rev | TTCGCAATGGTGCCGTCCAGC |
| 1A8-For | CTGCTGACCTGTGGCTTTGCT |
| 1A8-Rev | CCATTGAGCATCGGCGAAAT |
| 2B28-For | TCTTTTTGATCCCAATGACGCATT |
| β-Actin-For | CTGGCGGCACCACCATGTACCCT |
| β-Actin-Rev | GGAGGGGCCGACTCGTCATACT |
| ChIP assay | |
| 2B15-ChipF1 | TGCCGTTTGAGTTGTATAATTACTTCTTC |
| 2B15-ChipR1 | GAATAGGTGCATGCCAAGGAGACC |
| 2B15-ChipF2 | GGGGGGTTAGAGGCTATTGTATAGA |
| 2B15-ChipR2 | GAATATGTAAGTAACCTGTCTTATGT |
| 2B15-ChipR3 | TTAATATCGTGGTGCAAGTAATGTCTTC |
| 2B15-ChipF3 | GAGTGGGGGGTTAGAGGCTATTGTATAG |
| HCG3-ChipF | ATGGTGGACTACTACGAGGT |
| HCG3-ChipR | CACCTGCTTGAATCTCCTCT |
| TFF1/pS2-For | GGCCATCTCTCACTATGAATCACTTCTGC |
| TFF1/ps2-Rev | GGCAGGCTCTGTTTGCTTAAAGAGCG |