Global gene expression profiles of canine macrophages and canine mammary cancer cells grown as a co-culture in vitro

Table 1 Primers used for real-time qPCR

Gene symbol	Forward primer	Reverse primer	Optimum annealing temp. (°C)	Optimum annealing time (sec)
CCL2	CTCCAGTCACCTGCTGCTAT	CACAGCTTCTTTGGGACACT	60	4
CCL3	CCAGGTCTTCTCACCATTTG	AGATAATACCGGGCTTGGAG	60	5
CD163	ATGTCCAGTGTCCAAAAGGA	CATGTGATCCAGGTCTCCTC	61	6
CSF1R	TGCAGTTTGGGAAGACTCTC	TGTGGACTTCAGCATCTTCA	60	4
HIF1	GATTGCAGCTCCATCTCCTA	TCCTTTTCCTGCTCTGTTTG	58	5
IL18	GATATGCCCGATTCTGACTG	GCCTGGAACACTTCTCTGAA	60	9
MMP9	CGACTACGACCAGGACAAAC	AAGCCCCACTTCTTGTCTCT	61	8
VEGF-C	CAGCAACACTACCACAGTGC	CTCCAGAATTTGAGGCAAAA	61	5
Wnt7b	GCGGAGGGCTGTGTATAAGA	GTCCCCTACTTTGCGGAACT	59	5
HPRT	AGCTTGCTGGTGAAAAGGAC	TTATAGTCAAGGGCATATCC	59	6
RPS19	CCTTCCTCAAAAAGTCTGGG	GTTCTCATCGTAGGGAGCAAG	61	10

Primers sequences used in this study and their annealing optimal temperature and time. The mRNA sequences of key genes were obtained from NCBI database. Primers were designed using PRIMER3 software (free on-line access) and checked using Oligo Calculator (free on-line access) and Primer-Blast (NCBI database). Primers sequences are listed in Table 1. HPRT and RPS19 genes were used as non-regulated reference genes for normalization of target gene expression [3, 9]

ISSN: 1746-6148