Table 1.
Details of qPCR assays
| Gene name | GeneBank transcript Acc. Number | Primers’ sequences | qPCR reaction conditions | qPCR reaction content |
|---|---|---|---|---|
| IL-1α | NM_000575.4 |
5’-TAGGTCAGCACCTTTTAGCTTC 5’-GTATCTCAGGCATCTCCTTCAG |
95 °C, 3 min; 45x (95 °C, 5 s; 59 °C, 10 s; 72 °C, 10 s; 75 °C, 10 s—sample reading) Melting curve: 95 °C, 15 s; 60 °C, 1 min; 60 °C → 95 °C reading every 0.3 °C |
5 µl AmplifyMe NoRox SybrGreen (with SybrGreen fluorophore) (Blirt, Poland), 200 nM each primer, Σ 10 µl |
| IL-1β | NM_000576.2 |
5’-CCTTAGGGTAGTGCTAAGAGGA 5’-TACAGACACTGCTACTTCTTGC |
||
| IL-1RN | NM_173841.2 |
5’-GGCACTTGGAGACTTGTATGAA 5’-GAGCTGAAGTCACAGGAAGTAG |
||
| IL-6 | NM_000600.4 |
5’-CACTCACCTCTTCAGAACGAAT 5’-AGGCAAGTCTCCTCATTGAATC |
||
| INF- γ | NM_000619.2 |
5’-TGGAAAGAGGAGAGTGACAGAA 5’-TATTGCTTTGCGTTGGACATTC |
||
| TGF-β1 | NM_000660.6 |
5’-GAGCTGTACCAGAAATACAGCA 5’-AACTCCGGTGACATCAAAAGAT |
||
| GUSB | NM_000181.4 |
5’-ATGCAGGTGATGGAAGAAGTGGTG 5’-AGAGTTGCTCACAAAGGTCACAGG |