Table 5 Primers used for quantitative real-time RT-PCR. Primers were designed using NCBI’s Primer BLAST. Ten DEGs were selected to validate RNA-sequencing results by qRT-PCR. ACTB, GAPDH and 18S rRNA were used as reference genes. The designed primers are listed with product length and annealing temperature. F forward primer, R reverse primer
Gene | Primer sequence 5′-3’ | Product length (bp) | Annealing Temp C° | Accession no./reference of target transcript |
|---|---|---|---|---|
SLC10A2 | F: ATCGTTCACCTACGAGGAGC | 191 | 66 | XM_001493450.3 |
R: TCACCTTGTGGAGCGATGAC | Â | Â | Â | |
SLC16A9 | F: TGTTCTTTGCTGGGCTTGGA | 110 | 68 | XM_023643589.1 |
R: CAGGACGCAGAAGCCACTAA | Â | Â | Â | |
ITGB3 | F: GCACCCGTTACTGTCGTGAT | 145 | 65 | NM_001081802.1 |
R: AGGATGGACTTTCCACTGGC | Â | Â | Â | |
THBS2 | F: TGGCTGGAAAGACTACACCG | 107 | 65 | NM_001163117.2 |
R: CTGAATCCGCCATGACCTGT | Â | Â | Â | |
FN1 | F: GGTCGTTACTGTGGGCAACT | 101 | 65 | XM_023642280.1 |
R: CCTCTCCGATGGCGTAATGG | Â | Â | Â | |
COL6A1 | F: CCTCCTGGGATAAACGGCAC | 184 | 65 | XM_001488351.5 |
R: ACTCGTCCATCTCTGGTCGT | Â | Â | Â | |
ACKR3 | F: ATGCCTGAGTAGCCTGGAGA | 113 | 65 | XM_023642191.1 |
R: GTCCTGTGGTGATGCAAACG | Â | Â | Â | |
LOC100073089 (ENPP3) | F: TAGAATACGTGGTCAACACCAG | 190 | 68 | XM_023651094.1 |
R: TCAACCCAGTTGGCTTCCTG | Â | Â | Â | |
PDE10A | F: GCGTGAATTGTAGCAGCCAG | 76 | 65 | XM_023633145.1 |
R: ACTGATTGCAGAAAGACACTTCC | Â | Â | Â | |
MMP25 | F: ATGTCACCGTCAGCAACACAG | 189 | 70 | XM_023633145.1 |
R: GTCCAGGCTTGAGAGTGGCT | Â | Â | Â | |
ACTB | F: TCCCAGCACGATGAAGATCAA | 189 | 68 | XM_023655002.1 |
R: GGTGGATCGCACTAACAGT | Â | Â | Â | |
GAPDH | F: ATTGCCCTCAACGACCACTT | 140 | 70 | NM_001163856.1 |
R: TCTTGCTGGGTGATTGGTGG | Â | Â | Â | |
18S rRNA | F: GCGTGTGCCTACCCTACGCC | 165 | 68 | AJ311673.1/ [24] |
R: ATCGTTCACCTACGAGGAGC | Â | Â | Â |