FIG 3.

Generation of wild-type and D614G pseudoviruses for functional assays by Nb (Ty1)-CBD-functionalized cellulose. (a) Schematic overview of the pseudovirus production. HEK293T cells were transfected with a lentiviral vector expressing a green fluorescent protein (GFP), a plasmid encoding SARS-CoV-2 spike, and packaging vectors. The transfected cells produced lentiviral particles pseudotyped with the S protein of SARS-CoV-2, and the pseudovirus can transduce HEK293T expressing human angiotensin-converting enzyme 2 (hACE2) to express GFP. (b) Microscope images showing that the HEK293T-hACE2 cells expressed GFP after transduction with lentivirus pseudotyped with the wild-type (WT) SARS-CoV-2 spike protein or the D614G variant. Scale bar, 100 μm. (c) Representative flow cytometric analysis evaluating the transduction efficiency of SARS-CoV-2 WT and D614G pseudoviruses compared with two negative-control groups: HEK293T-hACE2 without any transduction and HEK293T transduced with SARS-CoV-2 WT pseudotyped lentivirus. Results are representative of three independent experiments.