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. 2022 Jun 8;221(7):e202108015. doi: 10.1083/jcb.202108015

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© 2022 Wang et al.

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Figure 7. — SARS-CoV-2 ORF10 infection impairs trachea cilia. (A) Schematic of the experimental design for the SARS-CoV-2 ORF10 infection system used in the present study. The dark pink lung symbols in rows two and three means virus infection. (B) ORF10 expression was detected in the trachea of hACE2 mice. Immunofluorescence analysis for HA (magenta) and FOXJ1 (white) was performed in Spike-lentivirus-ORF10 infected trachea. Nuclei were stained with DAPI (blue). The white arrowhead indicates FOXJ1 and ORF10 positive cells. (C) IFT46 was downregulated in mouse lungs after Spike-lentivirus-ORF10 infection. Immunoblotting analysis of IFT46, HA, and GFP was performed in mock, Spike-lentivirus, and Spike-lentivirus-ORF10 infected hACE2 mouse lungs. Tubulin served as a loading control. (D) Haematoxylin and eosin (H&E) staining of hACE2 lungs after mock, Spike-lentivirus (ORF10−), and Spike-lentivirus-ORF10 (ORF10+) infection. The black arrows indicate epithelial damage and the dashed lines indicate the boundary of the cilia layers. (E) ORF10 damaged the cilia layer in mouse trachea. Immunofluorescence analysis for Ac-Tubulin (magenta) and HA (white) was performed in mock, Spike-lentivirus, and Spike-lentivirus-ORF10 infected mice. Nuclei were stained with DAPI (blue). (F) Quantification ratio of ciliated epithelial cells in E (n = 5 independent experiments). Data are presented as the mean ± SEM. *** indicates P < 0.001. Statistical analysis was performed with two-tailed unpaired Student’s t test. Source data are available for this figure: SourceData F7.