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. 2021 May 18;10(1):894–904. doi: 10.1080/22221751.2021.1925163

Figure 1.

Figure 1.

Generation of SARS-CoV-2 pseudoviruses containing miRFP reporter. (A) Schematic drawing of plasmids expressing full-length (S) and truncated (Str) SARS-CoV-2 S proteins, truncated S proteins with mutations (D614G and AAAR), and VSV-G protein as well as co-transfection with pNL43 R-E-miRFP (miRFP) to generate different pseudoviruses containing miRFP reporter, infection of target cells, one-step imagining and various applications. (B) Regular and spin infections of VSV pseudovirus to HEK-293T-hACE2 cells with miRFP signals at 48 h post-infection (top) and infection kinectics at 24–96 h post-infection (bottom). Data are means and standard deviations of triplicates from one representative experiment of two. (C) Yields of SARS-CoV-2 pseudovirus Str determined in HEK-293T-hACE2 cells at 48 h post-infection comparing different ratios of plasmid to vector (4, 2, 1, 0.5 µg/2 µg) during transfection, media, miRFP vector only and VSV pseudovirus (2 µg/2 µg) as positive control. miRFP signals (top) and quantification (bottom). **** P < 0.0001, *** P < 0.001, ** P < 0.01 by two-way (B) and one-way (C) ANONA (GraphPad 6.0).