Fig. 6.

Statin alters ACE2 localization in SARS-CoV-2 infected airway organoids. (A) The scheme of Statin analogs treatment to the SARS-CoV-2 infected AO. (B) We used immunoblotting analysis and (C) immunofluorescence staining to assay the inhibition effect on the N protein production in SARS-CoV-2-infected AO after 10 µM Fluvastatin or Simvastatin treatment. We detected the ACE2, TMPRSS2, and N protein levels in the indicated viruses/Statin combinations. GAPDH protein was used as a loading control. Immunofluorescence staining of N protein in the AO. The AO cells were stained for N protein (green) or EpCAM (red) and counterstained with DAPI (blue). Scale bar = 10 μm. (D, E) We used immunofluorescence staining to trace the ACE2 internalization process after indicated concentration of Fluvastatin or Simvastatin treatment. The airway organoid cells were stained for ACE2 (green) and counterstained with DAPI (blue). Scale bar = 10 μm. (F) Geographical drawing of amino acid changes in the spike protein of Wuhan, Alpha, Delta, and Omicron SARS-CoV-2 variants. The differences are shown in reference to SARS-CoV-2 Wuhan-1 genome, labeled with triangle. NTD, N-terminal domain; RBD, receptor binding domain; RBM, receptor binding motif; SD1, subdomain 1; SD2, subdomain 2; N, nucleocapsid. (G) Luciferase assay for quantification of SARS-CoV-2 pseudovirus infection (one MOI) in the 10 µM Fluvastatin or Simvastatin treated HEK293-ACE2 cells. Until 48 h later, we harvested the cell lysate for detecting the luciferase activity. The luciferase activity of each sample was normalized to the untreated control as 100%. Significance was determined by two-way ANOVA (*, p < 0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)