Oligonucleotides used in this study These oligonucleotides were used to amplify UGT1A7, UGT1A8, or UGT1A10 promoters, to mutate the HNF4α response element in the UGT1A9 promoter, and for EMSA or ChIP analyses. Underlined nucleotides represent restriction sites used to clone the genomic PCR fragments, whereas bases in boldface type correspond to the HNF4α response element, and nucleotides in italic correspond to mutated bases. The apo CIII probe derives from Mietus-Snyder et al. (1992).
| Promoter cloning | |
| UGT1A7p (Xho I) sense | 5′-CTAGCACTCGAGCGAGACCAGCCTGG |
| UGT1A7p (HindIII) antisense | 5′-CTAGCTGAAGCTTATCAGAGAACTTCAGCCC |
| UGT1AS/10p (XhoI) sense | 5′-CTAGCACTCGAGCAGGGTTGTCAATGTCATTTC |
| UGT1A8p (HindIII) antisense | 5′-CTAGCTGAAGCTTATGAGAGAACTGCAGCCC |
| UGT1A10p (HindIII) antisense | 5′-CTAGCTGAAGCTTATGAGAGAACTGCAGCCC |
| Site-directed mutagenesis | |
| 1A9p1mt and 1A9p0.5mt | 5′-TTTGCTCTGGGACGGGCCTTGAAAAAAATTAG |
| 1A9pmt to 1A8 | 5′-TTGCTCTGGGATGAATTCCAAAAAA |
| 1A8pmt to 1A9 | 5′-TTGCTTTGGGACAAATTCCAAAAAT |
| EMSA | |
| 1A9HNF4α REwt | 5′-TTTGCTCTGGGACAAATTCCAAAAAAAATTAG |
| 1A9HNF4α REmt | 5′-TTTGCTCTGGGACGGGCCTTGAAAAAAATTAG |
| 1A7 –368/–355 | 5′-TTTTCTTTGTGACAAATTCCAAAATTATTAGG |
| 1A8 –364/–371 | 5′-TTTGCTTTGGGATGAATTCCAAAAATATTAGC |
| 1A10 –369/–356 | 5′-TTTGCTTTGGGATAAATTCCAAAAATATTAGC |
| Apo CIII | 5′-CAGCAGGTGACCTTTGCCCAGCGCCC |
| ChIP | |
| UGT1A9 –386 (sense) | 5′-TGAGTTGCCATCTTCTCTGG |
| UGT1A9 –241 (antisense) | 5′-ATGCTTTTGGACCTTGAAGGT |
| UGT1A9 –2050 (sense) | 5′-GATTACAGGCATGCACCACCACCT |
| UGT1A9 –1890 (antisense) | 5′-CTCACACCTGTAGTCCCAGCAC |
| β-actin forward | 5′-CGAGCCATAAAAGGCAACTTTCG |
| β-actin reverse | 5′-AGGAAGAGGAGGAGGGAGAGTTT |