Extended Data Fig. 3: Entry and replication characteristics of Omicron subvariant BA.5.

(a-d) Calu-3 cells were infected with 2000 E copies/cell at 37 °C or 32 °C, one representative of two independent experiments shown. (a) Viral replication by RT-qPCR and (b) infection levels by flow cytometry at 24hpi. (c) IFNB and (d) CXCL10 expression in cells from (a). (e) IFNB and CXCL10 expression in response to poly(I:C) transfection in Calu-3 cells at 24h of stimulation, two replicates shown. (f-j) Primary bronchial human airway epithelial cells (HAEs) were infected with 1500 E copies/cell of the indicated variants at 37 °C or 32 °C. Three biological replicates shown. Viral replication was measured by (f) intracellular E copies at 72hpi and viral release of (g) BA.2, (h) BA.4 and (i) BA.5 into apical washes over time. Relative expression of (j) IFNB and CXCL10 normalized to GAPDH in cells from (f). Fold changes are normalized to mock. Pairwise comparisons were performed using an unpaired two-tailed Student’s t-Test as indicated. For g-i, two-way ANOVA with a Bonferroni post-test was used to compare temperatures at each time point. Fold change over mock is shown. Mean + /-SEM or individual datapoints are shown. hpi, hours post infection.

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