Table 1.
Primer Sequences Used for RT-PCR Analysis of Gene Expression in Retrorsine-Exposed Rat Hepatocytes and SHPCs
| Primer name | Sequence (5′-3′) | Annealing temp. (°C) | Amplicon size (bp) |
|---|---|---|---|
| CYP reductase | GAGCACTTCAATGCCATGGGCAAG | 65 | 597 |
| GATGTCCAGGTAGTAGGTGAGGGCC | |||
| CYP 1A1 | GATGCTGAGGACCAGGAAGACCGC | 65 | 679 |
| CAGGAGGCTGGACGAGAATGC | |||
| CYP 1A2 | CTTGGAGAAGCGTGGCCAGG | 55 | 664 |
| CTACAAAGACAACGGTGGTCT | |||
| CYP 2B1/2* | CCAAGCCGTCCACGAGACTT | 60 | 380/404* |
| TTGGGAAGCAGGTACCCTC | |||
| CYP 2E1 | CTGCCCCCAGGACCTTTTCCC | 65 | 847 |
| ATCAGGAGCCCATATCTCAGAGTTGTCCTGGTGGTCTC | |||
| CYP 3A1 | CAGCTCTCACACTGGAAACCTGGG | 60 | 689 |
| CTCATATATTTGGGGTGAGGAATGG | |||
| CYP 4A3 | TCGAGGATCTAAACAACCTGAC | 60 | 573 |
| GGTTGTGATACCTTTGGGTATGG | |||
| C/EBP-α | TTCCAGATCGCACACTGCCC | 65 | 404 |
| TGACCAAGGAGCTCTCAGGC | |||
| C/EBP-β | TGACCAAGGAGCTCTCAGGC | 65 | 419 |
| ACCCACCACCACACACAACCC | |||
| C/EBP-γ | CACTGGACAGCTGGTTAGCC | 60 | 399 |
| TCTCGACGTGACCTGATGGG | |||
| HNF-1α | AGACACCAACCTCAGCACCC | 65 | 463 |
| AGACACCTCAGAGCCATCCC | |||
| HNF-1β | CCGTTCTTGGAAAATGGTGTCCAAGC | 60 | 235 |
| ATGGGAGGAGTGTCATAGTCGTCG | |||
| HNF-3α | TGCAGTACTCTCCTTACGGC | 60 | 714 |
| GGGGAAACTTGTTTAGCTGGG | |||
| HNF-3β | GCCCAGTCACGAACAAAGCC | 60 | 686 |
| GGAACTCTGGCATTCTAGCC | |||
| HNF-3γ | AACTGAAGTTGGATGCGCCC | 60 | 476 |
| ATATGTCCTCCGACATCGCG | |||
| HNF-4 | TCTACAGAGCATTACCTGGC | 60 | 614 |
| TGAGGGGAAGATGAAGACGG | |||
| HNF-6 | GACAAATGGCAGGACGAGGG | 60 | 681 |
| AGCGTACTGGTTTAGGTGCC | |||
| WT1 | GTCAGAAAAAGTTTGCGCGG | 55 | 200 |
| CCATCATGTCCTTTTGTAGG | |||
| α-Fetoprotein | CAGTGAGGAGAAACGGTCCG | 60 | 252 |
| ATGGTCTGTAGGGCTCGGCC | |||
| α-1-Antitrypsin | ATGGATTACCTGGGCAACGC | 60 | 398 |
| TTTTCCCACAAAGAGGGGGC | |||
| Albumin | AAGGCACCCCGATTACTCCG | 60 | 608 |
| TGCGAAGTCACCCATCACCG | |||
| Transferrin | CCTGACAAAACGGTCAAATGGTGC | 60 | 251 |
| TAAAAACTCTGCTGCCACAGGC | |||
| Tyrosine aminotransferase (TAT) | TACTCAGTTCTGCTGGAGCC | 65 | 471 |
| GCAAAGTCTCTAGAGAGGCC |
*The cytochrome P450 mRNA isoforms 2B1 and 2B2 have substantial homology, differing mainly in an additional 24 bp in 2B2. Therefore, these primers amplify both isoforms and amplicons can be distinguished by size on agarose gels.