The gene expression profiles of canine mammary cancer cells grown with carcinoma-associated fibroblasts (CAFs) 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)
DSP	CAGACTCACCGAAGAGGAAA	CTGCTGTGAAGTCTGGGAGT	61	7
MAG	TGCCATCGTCTGCTACATTA	CAGTCGCCTCTCACTCTCAT	60	6
PCDH19	CTTTCACATCACTGCACTCG	GTGTGTTGGGAGGTGAGTTC	61	6
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). HPRT and RPS19 genes were used as non-regulated reference genes for normalization of target gene expression [19, 20].

ISSN: 1746-6148