Author: Suddala, Krishna C.; Lee, Christine C.; Meraner, Paul; Marin, Mariana; Markosyan, Ruben M.; Desai, Tanay M.; Cohen, Fredric S.; Brass, Abraham L.; Melikyan, Gregory B.
Title: Interferon-induced transmembrane protein 3 blocks fusion of sensitive but not resistant viruses by partitioning into virus-carrying endosomes Document date: 2019_1_14
ID: 15wxk8lt_57
Snippet: For intraviral IFITM3 pseudovirus production, HEK293T/17 cells grown in 100-mm dishes were transfected with 2 μg of pCAGGS H1N1 HA/NA, 3 μg pR9ΔEnv, 1.5 μg BlaM-Vpr, 0.5 μg pcRev, and 5 μg of either empty Vector pQCXIP, IFITM3, or 2M-IFITM3 using JetPRIME reagent. The viral supernatants cleared of cellular debris as described above, were concentrated 10x, using Lenti-X Concentrator (Clontech, Mountain View, CA). Following overnight concentr.....
Document: For intraviral IFITM3 pseudovirus production, HEK293T/17 cells grown in 100-mm dishes were transfected with 2 μg of pCAGGS H1N1 HA/NA, 3 μg pR9ΔEnv, 1.5 μg BlaM-Vpr, 0.5 μg pcRev, and 5 μg of either empty Vector pQCXIP, IFITM3, or 2M-IFITM3 using JetPRIME reagent. The viral supernatants cleared of cellular debris as described above, were concentrated 10x, using Lenti-X Concentrator (Clontech, Mountain View, CA). Following overnight concentration with Lenti-X, virus was precipitated by centrifuging at 1439xg for 45 min, 4˚C, resuspended in DMEM without phenol red or FBS, and stored at -80˚C.
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