Fig. 7: Membrane-tethered mucins restrict SARS-CoV-2 Spike-mediated entry. | Nature Genetics

Fig. 7: Membrane-tethered mucins restrict SARS-CoV-2 Spike-mediated entry.

From: Genome-wide bidirectional CRISPR screens identify mucins as host factors modulating SARS-CoV-2 infection

Fig. 7

a, Schematic of the pseudotype entry assay using a VSV-CoV-2-S encoding GFP. Following infection, cells are imaged regularly over 24 h to track viral infection by quantifying GFP+ cell counts. hpi, hours postinfection. b, Time course of VSV-CoV-2-S infection of NTG, MUC1, MUC4, MUC21 and CD44-overexpressing GOF cell lines. The significance of the difference relative to NTG at 24 h postinfection was determined by two-sided t-test, n = 3 biological replicates. c., Time course of VSV-CoV-2-S infection of NTG and MUC4 GOF cells pretreated with StcE mucinase before VSV-CoV-2-S infection. Significance of the difference between StcE-treated cells versus untreated cells for each cell line at the point of saturation at 8 h postinfection was determined by two-sided t-test, n = 3 biological replicates. d, Time course of VSV-RABV-G infection in NTG and MUC4 GOF cells pretreated with StcE before pseudotype virus infection. VSV-RABV-G, VSV particles pseudotyped with rabies virus glycoprotein. Significance comparing StcE-treated versus untreated cells at 5 h postinfection was determined by two-sided t-test, n = 3 biological replicates. e, Pseudotype entry assay measuring GFP+ cells at 8 h postinfection comparing infection rates between NTG or mucin-overexpressing cells with and without StcE pretreatment. The full dataset is displayed in Extended Data Fig. 9. n = 5 biological replicates; two-tailed t-tests were performed comparing indicated pairs. The dotted line indicates the NTG average for two separate guides, with and without StcE treatment. f, Schematic depicting a SARS-CoV-2 binding assay. g, RT–qPCR of relative levels of viral N gene copies following a virus binding assay conducted on Calu-3 cells at an MOI of 2.5 with or without StcE treatment. Significance comparing StcE-treated cells versus untreated cells was determined by two-sided t-test, n = 6 biological replicates. Error bars denote mean ± s.e.m. NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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