Extended Data Fig. 6: Cytopathogenicity of SARS-CoV-2 recombinant viruses in nasal epithelium at day 4.
From: Phenotypic evolution of SARS-CoV-2 spike during the COVID-19 pandemic
a, Quantification of cell sloughing from hNECs infected with another clone of spike recombinant virus (n = 3) as in Fig. 4g, h, using whole-section software assisted imaging as described in Methods. b, Quantification of caspase-3 cleavage in hNECs (n = 3). Apoptotic cells were quantified using whole-section software assisted image analysis. c, Microphotographs showing apoptotic cells, as revealed by TUNEL assays, in hNEC infected with the indicated viruses at 4 dpi. Apoptotic nuclei are stained in brown. d, Quantification of apoptotic nuclei using whole-section software assisted image analysis as described in Methods. Three biological replicates are shown from two independent sets of clones for each spike recombinant virus. e, Microphotographs of hNEC infected with either S:B.1, S:BA.1 or S:EG.5.1 and stained with haematoxylin and eosin to reveal sloughing. f, Quantification of cell sloughing in infected hNEC cultures (n = 3). g, Microphotographs showing RNA in situ hybridisation for the detection of IFIT1 at 4 dpi in hNEC infected with the indicated viruses. h Quantification of IFIT1 expression of two independent sets of clones (each n = 3) using whole-section software assisted image analysis. In (a,b,d,f and h) data are mean ± standard error to the mean (SEM) of three biological replicates. Statistical significance between S:B.1 and other spike recombinant SARS-CoV-2 was determined by one-way ANOVA with multiple comparisons between conditions using the Holm-Ĺ Ădák post-hoc test. The calculated p-values are indicated in the figures. Scale bar: 200 ÎĽm.
