Author: 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
Title: Avian Interferon-Inducible Transmembrane Protein Family Effectively Restricts Avian Tembusu Virus Infection Document date: 2017_4_20
ID: 0pjg25kn_13
Snippet: were generated using lentiviral vectors as previously described . Briefly, 293T cells were cotransfected with shRNA construct and HIV-based packaging constructs (pLP, pLP1, and pLR2). Supernatant of the cultured cells containing pseudotyped lentiviruses with indicated shRNAs were collected at 48 h post-transfection and filtered through the 0.22 µM syringe-driven filter. DF-1 single-cell suspension was incubated with the supernatant and 8 µg/mL .....
Document: were generated using lentiviral vectors as previously described . Briefly, 293T cells were cotransfected with shRNA construct and HIV-based packaging constructs (pLP, pLP1, and pLR2). Supernatant of the cultured cells containing pseudotyped lentiviruses with indicated shRNAs were collected at 48 h post-transfection and filtered through the 0.22 µM syringe-driven filter. DF-1 single-cell suspension was incubated with the supernatant and 8 µg/mL of polybrene (Sigma) and centrifuged at 2,100 rpm, 32 • C for 120 min. DF-1 cells were then cultured in DMEM supplemented with 10% FBS for further studies. qRT-PCR was performed to determine the interference efficiency and mRNA expression of viral genes in DF-1 cell lines at indicated times after infection. The viral titers of supernatants were examined by TCID 50 assay in DEF cells.
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