Avian Interferon-Inducible Transmembrane Protein Family Effectively Restricts Avian Tembusu Virus Infection

Chen, Shilong; Wang, Long; Chen, Jieying; Zhang, Lanlan; Wang, Song; Goraya, Mohsan U.; Chi, Xiaojuan; Na, Yang; Shao, Wenhan; Yang, Zhou; Zeng, Xiancheng; Chen, Shaoying; Chen, Ji-Long

Quantitative Real-Time RT-PCR and RT-PCR Total RNA was extracted from the cultured cells and duckling spleen tissues using Trizol reagent (TransGen Biotech, Beijing, China) according to the manufacturer's instructions. Equal amount of RNA (4 µg) was reverse transcribed into cDNA utilizing M-MLV Reverse Transcriptase (Promega, USA). The cDNA was analyzed by qRT-PCR using TransStart Green qPCR SuperMix (TransGen) and PCR using rTaq DNA polymerase (Takara Bio). The primers specific for chicken β-actin, IFNa, IFN-β, IFN-λ, and ATMUV E gene have been previously described (Chen et al., 2016a) . The primers specific for ATMUV NS5, duck β-actin, IFN-a, IFN-β, IFN-λ, IFITM1, 2, 3, and chicken IFITM1, 2, 3 were designed using the Primer5 software ( Table 1) . The relative mRNA abundances were analyzed using the 2 ′ Ct method with housekeeping gene (β-actin) as an internal normalization and plotted as fold changes compared with the mock-infected samples.