Author: Wilkins, Jordan; Zheng, Yi-Min; Yu, Jingyou; Liang, Chen; Liu, Shan-Lu
Title: Nonhuman Primate IFITM Proteins Are Potent Inhibitors of HIV and SIV Document date: 2016_6_3
ID: 1m1woscb_10
Snippet: SupT1 and Jurkat-inGLuc cell lines stably expressing IFITMs were produced by transduction of cells with pQCXIP-IFITM constructs [19, 29] . Briefly, HEK293T cells were seeded in 35 mm dishes and transfected with 1 μg pQCXIP-flag-IFITM, 1 μg pCMV-gag-pol-MLV, and 0.1 μg pVSV-G using calcium phosphate method. Viral supernatants were harvested over 72 hours and clarified via centrifugation. Retroviral particles were used to infect SupT1 or Jurkat-.....
Document: SupT1 and Jurkat-inGLuc cell lines stably expressing IFITMs were produced by transduction of cells with pQCXIP-IFITM constructs [19, 29] . Briefly, HEK293T cells were seeded in 35 mm dishes and transfected with 1 μg pQCXIP-flag-IFITM, 1 μg pCMV-gag-pol-MLV, and 0.1 μg pVSV-G using calcium phosphate method. Viral supernatants were harvested over 72 hours and clarified via centrifugation. Retroviral particles were used to infect SupT1 or Jurkat-inGLuc cells in the presence of 4 μg/ml polybrene by spinoculation at 1,850 x g for 1 hour at 4˚C. Stably transduced cells were selected for with 1 μg/ml puromycin.
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