Quantitative PCR Analysis of PIR2 Expression

RNA extraction and cDNA synthesis were done as described before^{59 (link)}. Equal amount of RNA was used in cDNA synthesis. Quantitative PCRs were performed with TaqMan Universal Master mix with MJ Research DNA Engine Opticon2 or with QuantStudio-3 Real-Time PCR system. All TaqMan assays were purchased from Applied Biosystems. For semi-quantitative PCR the quality of cDNA was tested by GAPDH amplification (GF: 5′-GGCTGAGAACGGGAAGCTTGTCAT-3′ and GR: 5′-CAGCCTTCTCCATGGTGGTGAAGA-3′). PIR2 expression was analysed with the primers: FL-PIR2-F: 5′-ATGGGCTCAGCTGGTAGGC-3′ and FL-PIR2-R: 5′-GGTTGTGGATGGGTCGTGCT-3′. The amplified DNA fragments were analysed as described previously^{60 (link)}.