Extended Data Fig. 5: Validation of syncytium-enhancing mutations at the furin cleavage site region in D614G Spike. | Nature Biomedical Engineering

Extended Data Fig. 5: Validation of syncytium-enhancing mutations at the furin cleavage site region in D614G Spike.

From: High-throughput screening of genetic and cellular drivers of syncytium formation induced by the spike protein of SARS-CoV-2

Extended Data Fig. 5: Validation of syncytium-enhancing mutations at the furin cleavage site region in D614G Spike.The alternative text for this image may have been generated using AI.

a) GFP-split complementation system was applied for assessing the syncytium-enhancing mutations at the furin cleavage site region in D614G Spike. A549-ACE2-GFP1-10 receiver cells and HEK293T-GFP-11 sender cells that express WT D614G Spike were used. Representative images are shown. b) Quantification of the syncytium area and average size of syncytium for Spike variants in (a). Data shown are mean ± SD (n = 9). P-values indicated were compared with the D614G WT. c-d) Cell surface staining of Spike furin cleavage site variants in HEK293T sender cells. Anti-Spike antibodies against its S2 and S1 subunit were used to evaluate the total surface expression of Spike protein and the level of S1-cleaved Spike on cell surface, respectively. Ctr represents the control cells without Spike expression. In (d), the TMPRSS2 and Furin groups are HEK293T sender cells overexpressed with TMPRSS2 and Furin, respectively. HEK293T sender cells used in (c) and the ‘none’ group in (d) express only minimal levels of TMPRSS2 and Furin. The percentage of Spike cleavage is determined by the proportion of S1-stained cells. Data shown are mean ± SD (n = 3). P-values indicated were compared with the D614G WT in each group (that is, Ctr/TMPRSS2/Furin). Statistical significance was determined using one-way ANOVA. ns: no significance. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. n indicates the number of biological replicates.

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