Author: Shi, Chong-Shan; Nabar, Neel R.; Huang, Ning-Na; Kehrl, John H.
Title: SARS-Coronavirus Open Reading Frame-8b triggers intracellular stress pathways and activates NLRP3 inflammasomes Document date: 2019_6_5
ID: 0fpa1f30_33
Snippet: HEK293 cells were seeded onto 24-well plates in standard media. The following day, the media was changed to OPTI-MEM (Gibco) and the cells transfected with constructs expressing NLRP3 Flag (15 ng), Casp-1 Flag (5 ng), ASC (5 ng), full-length IL-1β Flag (150 ng) and either a control plasmid (GFP or Flag) or GFP-8b or 8b-Flag. Cell supernatants and lysates were immunoblotted for fulllength and mature IL-1β the next day......
Document: HEK293 cells were seeded onto 24-well plates in standard media. The following day, the media was changed to OPTI-MEM (Gibco) and the cells transfected with constructs expressing NLRP3 Flag (15 ng), Casp-1 Flag (5 ng), ASC (5 ng), full-length IL-1β Flag (150 ng) and either a control plasmid (GFP or Flag) or GFP-8b or 8b-Flag. Cell supernatants and lysates were immunoblotted for fulllength and mature IL-1β the next day.
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