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. 2021 Dec 2:eabj5508. doi: 10.1126/science.abj5508

Fig. 5. Efficacy of 4′-FlU against SARS-CoV-2 replication in HAE organoids.

Fig. 5.

(A) Multicycle growth curve of SARS-CoV-2 WA1 isolate on ALI HAE from two donors. Shed virus was harvested daily and titered by plaque assay (n = 3). (B and C) Confocal microscopy of ALI HAE cells from “F1” donor mock-infected (B) or infected (C) with SARS-CoV-2 WA1 isolate, at 3 days post-infection. SARS-CoV-2 infected cells, goblet cells, and nuclei were stained with anti-SARS-CoV-2 N immunostaining, anti-MUC5AC immunostaining, and Hoechst 34580, pseudo-colored in red, green, and blue, respectively. z-stacks of 35 μm slices (1 μm thick) with 63× objective with oil immersion. Dotted lines represent the location of x-z and y-z stacks; scale bar, 20 μm. In all panels, symbols represent independent biological repeats and lines represent means. (D) Virus yield reduction of SARS-CoV-2 WA1 clinical isolate shed from the apical side in ALI HAE after incubation with serial dilutions of 4′-FlU on the basal side (n = 3). (E and F) Confocal microscopy of ALI HAE cells infected with SARS-CoV-2 WA1 isolate, and treated with 50 μM 4′-FlU 3 days after infection. Rare ciliated cells positive for N are represented in (F).