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. 2023 May 22;92:104608. doi: 10.1016/j.ebiom.2023.104608

Fig. 4.

Fig. 4

Hyperacute SARS-CoV-2 infection of the bronchiolar epithelium in a COVID-19 patient. Confocal and brightfield images of sections through lung tissue of COVID 21, who died two days after diagnosis of infection. The extracellular matrix in the bronchiolar wall emits a weak green or red autofluorescent signal. (a) N puncta reveal a patchy pattern of cells in the bronchiolar epithelium, and the dense packing of N puncta reflects high levels of intracellular viral RNA. The dashed square in (a) is magnified in (b). Most cells harbouring N puncta also harbour perinuclear S-sense puncta, reflecting ongoing viral replication. (c) S puncta are densely packed in many cells, and some of these harbour perinuclear orf1ab-sense puncta. (d) The bronchiole in (a) is shown in a brightfield image of a nearby H&E-stained section. There is marked atypia without necrosis in the bronchiolar epithelium and the surrounding alveoli are not affected. (e) M puncta are densely packed in ciliated cells harbouring FOXJ1 puncta, and some of these harbour perinuclear S-sense puncta. (f) Nucleocapsid-immunoreactive signal diffusely fills two ciliated cells harbouring FOXJ1 puncta and perinuclear N-sense puncta. (g) Basal cells containing KRT5/6-immunoreactive signal do not harbour orf1ab-sense puncta; by contrast, orf1ab-sense puncta cluster with FOXJ1 puncta in ciliated cells. (h) Club cells contain CC16-immunoreactive signal and do not harbour orf1ab puncta; by contrast, orf1ab puncta cluster with FOXJ1 puncta in ciliated cells. (i) MUC5AC-immunoreactive signal labels secreted mucus lining the mucosa; MUC5AC-immunoreactive goblet cells do not harbour N-sense puncta; a few cells harbouring FOXJ1 puncta also harbour N-sense puncta. (j) A clump of CD68-immunoreactive macrophages fills the bronchiolar lumen. The dashed square in (j) is magnified in (k). A few CD68-immunoreactive macrophages harbour S-sense puncta in areas indicated with a stippled line. (l) N puncta and S-sense puncta are not detected in the alveoli. DAPI served as nuclear stain. The names of RNAscope probes are in italics and the names of antigens/proteins that are revealed as immunoreactive signal by antibodies are in roman.