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Image Search Results
Journal: bioRxiv
Article Title: Isolation of ACE2-dependent and -independent sarbecoviruses from Chinese horseshoe bats
doi: 10.1101/2023.03.02.530738
Figure Lengend Snippet: ( A ) V976L mutation emerged in 190366 virus stocks. ( B ) Pseudotyped particles were produced with full-length spike WT or the V967L mutant. Spike was detected in producer cells and pseudotyped by western blotting for FLAG. ( C ) Indicated cells were infected with viral pseudotyped in the presence or absence of trypsin. ( D ) Schematic overview of the dual-reporter fusion assay developed for this study. T7 polymerase drives expression of GFP and luciferase separated by a P2A fusion peptide. ( E ) HEK 293T cells expressing receptor or, ( F) empty vector and T7-polymerase were combined with cells expressing spike and the T7-driven reporter. Luciferase was measured as a readout for cell fusion. Dotted lines indicate data from 1:4 ratio of receptor:spike cells. ( G ) Overview of 190366 spike with in silico predicted trypsin digest sites indicated. Location of V976L is indicated in green. ( H ) Concentrated viral pseudotyped were combined with a wide range or trypsin dilutions or ( I ) a fine range of trypsin dilutions and incubated at 37°C. Spike digestion was assessed by western blot for FLAG epitope.
Article Snippet:
Techniques: Mutagenesis, Produced, Western Blot, Infection, Single Vesicle Fusion Assay, Expressing, Luciferase, Plasmid Preparation, In Silico, Incubation, FLAG-tag