Table 1. Primers and probes used for gene expression analysis.
TagMan probe based qPCR assays according to Haarder et al. (2016). All the assays were optimized to have primer-annealing temperature at 60°C and amplification efficiencies of 100% ±5%. The three genes acbt2, eelf1a1l1 and rpl13a served as reference genes. All the nucleotides are oriented from the 3’end towards the 5’end. The probes were labelled with FAM at 3’end and with BQ1 at the 5’end. Genes names according to ZFIN.org.
| Gene | Forward primer | Reverse primer | Probe | GenBank Acc.no. |
|---|---|---|---|---|
| acbt2 | CCATCCTTCTTGGGTATGGA | ACAGGTCCTTACGGATGTCG | TGCGGTATCCACGAGACCACC | BC154531 |
| cd4-1 | CCAGACTGGAGCCAAGAGAC | CAGTGCAGCTCCACATCACT | CCTGCGCACAGTCAGGGAAA | EF601915 |
| cd8a | TCGGAGGTTGTGGACTTTTC | TAATGGTGGGGACATCGTCT | CGTTCTGCTGATCGCAACCA | BC114236 |
| cxcl8a | GATCTGTCTGGACCCCTCTG | GGGCATTCATGGTTTTCTGT | CCATGGGTTAAGAAGATCATTGATAGG | XM_001342570 |
| eelf1a1l1 | GAACGACCCACCCATGGAGG | TGATGACCTGAGCGTTGAAG | GAACGACCCACCCATGGAGG | AY422992 |
| gata3 | TACGTGTCCCGCTTAAAACC | TGAAGGGGCAATGAAGAAAG | GGCCTTCACTTTCGCCTGCT | BC162389 |
| Ifng1 | TATGGGCGATCAAGGAAAAC | CTTTAGCCTGCCGTCTCTTG | CGATCGTCCAGCGAAAGGCT | AB158361 |
| ighm | TGCAGTTCTGGTTCTGATGG | TGCACAAAATCGCTCAAATC | AATCACCCTCGGCTGCTTGG | AY643753 |
| ighz | ATTGGATGTCTGGCCTCTGA | AATGCTGGGTGACGTTTTTC | TGCACAAAATCGCTCAAATC | AY643750 |
| il1b | CGCTCCACATCTCGTACTCA | ATACGCGGTGCTGATAAACC | GAAGGAGACCGGCAGCTCCA | BC098597 |
| Il4 | AACTCTCTGCCAAGCAGGAA | AAACGCTGCAGTTTCCAGTC | GAGACAGCTGAATGCTTATGCAGCA | AB375405 |
| il 6 | AGACCGCTGCCTGTCTAAAA | CAACTTCTCCAGCGTGATGA | TCCGCATGGACTCGCAAGAC | JN698962 |
| il10 | CTTGCCAAAATCCCTTTGAA | ATCAAGCTCCCCCATAGCTT | TGAAAAGATGAAGGAAAAGGGGG | BC163038 |
| il12a | AGCAGGACTTGTTTGCTGGT | TCCACTGCGCTGAAGTTAGA | TAACTCGTCCTGCTCGGCCC | AB183001 |
| Il13 | GTCAGGCTGAGGAGGAGATG | AGCAGCGTGACTCCTGATCT | CTGGCCTGTCCGGTGTCAAA | AB375404 |
| il17a/f1 | GCTTTCTTATGGTGGCTTGC | GCCGGTATGAATGATCTGCT | ATCATTCGGTGCTGAGGGGG | AB195256 |
| il17a/f2 | TCAATCTGAGGACGGAAAGG | GCTCCATCTCCTGTTTCAGC | CATCTTGCCCACTGGTGTGGA | NM_001020798 |
| il17a/f3 | GGCTGCACATGTGTTTTACC | CAATGTGATTCGTTTTCAGGCT | CAGCGCCTATAGAAATAATCACTCCG | AB195260 |
| il 22 | GGATTACGCCAAAGGTGAAA | CGAGCACAGCAAAGCAATAA | CGACATCGAGGAACAACGGTG | NM_001020792 |
| mmp9 | ACAGGGAGACGCTCATTTTG | TGTTCCCTCAAACAGGAAGG | CGGTAATGCTGAGGGTGCAATGTGT | BC160656 |
| rpl13a | TCCCAGCTGCTCTCAAGATT | ACTTCCAGCCAACTTCATGG | CACACGCAAATTTGCCCTGC | NM_212784 |
| s100a10b | GCAGAGGGGAACTCATCAAC | CCCACCACAAGAGACACAAA | CAGCTGATGGAGAAAGAGCTGGC | BC163954 |
| saa | CTTGCTGTGCTGGTGATGTT | CTTCCAATTGGCCTCTTTCA | CGCTGGAGGTGCAAAGGACA | BC081487 |
| tbx21 | AAATCCAGGAGCATGGACAG | TGAGACTGGATGTGGGTTTG | TGGTTTTCCTATTGGAAGGCGG | AM942761 |
| tgfba | TGCGCAAGCTTTACATTGAC | AGGACCCCATGCAGTAGTTG | TGGATCCACAAGCCCAAGGG | AY178450 |
| tnfa | GCGCTTTTCTGAATCCTACG | AAGTGCTGTGGTCGTGTCTG | TGCACGCAGGAGCCTGAATC | AY427649 |