Table 1. Primer sequences and qRT-PCR conditions for determination of cytokine expression in the spleens of dogs with visceral leishmaniasis.
Primer | Forward primer (5′ – 3′) | Reverse primer (5′ – 3′) | Genbank reference sequence | Annealing temperature |
18S | CACGGCCGGTACAGTGAAAC | CCCGTCGGCATGTATTAGCT | M27358 | 60° |
CCL19 | GTAGACTGCCTGCCGTTGTGTTCA | ACTGCTGTGGCCCTTGTTCTTTG | NM_001005256.1 | 60° |
CCL21 | GCACAGGACTGTTGCCTCAA | CGGTAGCTGCGGACAACCT | NM_001005258 | 55° |
CCR7 | GGTGGTGGCTCTCCTTGTCATTTT | AGTTCCGCACGTCTTTCTTGAAGC | XM_548131.2 | 55° |
CXCL13 | GGGTGCCCAAAAAGAGAAATC | GATGGGAGGGTTCAAGCATACA | XM_845089.1 | 55° |
LT-α | GAGACCCCAGCATCCAGAAC | AAGGCACGATCCGTGTTTG | XM_843793.1 | 60° |
LT-β | CGGCTGGGAGGCGAAGAAAG | GGTAGGCGACGTGACAGTAGAGGT | NM_001033510.1 | 60° |
IL10 | CCACGACCCAGACATCAAGAA | ACAGGGAAGAAATCGGTGACA | U33843 | 60° |
TGF-β | CCACTGTTCCTGTGACAGCAA | GTCGGTTCATGCCATGAATG | L34956 | 60° |
TNF-α | AGCAAACCCCGAAGCTGAG | CGGCACTATCAGCTGGTTGTC | S74068 | 60° |
IFN-γ | AAGGAAGACATGCTTGGCAAG | CCTGCAGATCGTTCACAGGAA | NM_001003174 | 60° |