PMC:7100515 / 38744-39718 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/7100515","sourcedb":"PMC","sourceid":"7100515","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7100515","text":"Production of SARS-CoV-2 S pseudovirions and virus entry\nPseudovirions were produced by co-transfection 293T cells with psPAX2, pLenti-GFP, and plasmids encoding either SARS-CoV-2 S, SARS-CoV S, VSV-G, or empty vector by using polyetherimide (PEI). The supernatants were harvested at 40, 64 h post transfection, passed through 0.45 μm filter, and centrifuged at 800 × g for 5 min to remove cell debris. To transduce cells with pseudovirions, cells were seeded into 24-well plates and inoculated with 500 μl media containing pseudovirions. After overnight incubation, cells were fed with fresh media. About 40 h post inoculation, cells were lysed with 120 μl medium containing 50% Steady-glo (promega) at room temperature for 5 min. The transduction efficiency was measured by quantification of the luciferase activity using a Modulus II microplate reader (Turner Biosystems, Sunnyvale, CA, USA). All experiments were done in triplicates, and repeated at least twice or 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