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. 2022 Jun 16;13(4):e00519-22. doi: 10.1128/mbio.00519-22

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Copyright © 2022 Yamamoto et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 6 — The metalloproteinase-dependent entry pathway of authentic SARS-CoV-2 is involved in syncytium formation and cytopathicity. The effects of the drugs on the cytoplasmic viral RNA after SARS-CoV-2 infection are shown. Cells were treated with inhibitors for 1 h and added with SARS-CoV-2 at an MOI of 0.01 for HEC50B and HEC50B-TMPRSS2 cells and at an MOI of 0.1 for VeroE6, Calu-3, and A704 cells. The relative amount of viral RNA in the cells was normalized to cellular Rpl13a mRNA expression. Values are means ± SD (n = 3/group in panels a to d, n = 10/group in panel e). *, P < 0.05; **, P < 0.01. (a) Effects of marimastat or prinomastat on SARS-CoV-2 infection in HEC50B, A704, and VeroE6 cells. (b) Effects of marimastat and the inhibitor of the endosomal pathway on SARS-CoV-2 infection in HEC50B, A704, and VeroE6 cells. marima, 1 μM marimastat; E-64d, 25 μM E-64d; NH4Cl, 10 mM NH₄Cl. (c) Effect of marimastat, E-64d, and nafamostat on SARS-CoV-2 infection in HEC50B-TMPRSS2 cells. marima, 1 μM marimastat; E-64d, 25 μM E-64d; nafamo, 10 μM nafamostat. (d) Effects of selective metalloproteinase inhibitors on SARS-CoV-2 infection in HEC50B cells. GW, GW280264X; GI, GI254023X. (e) Effect of ADAM10 knockdown on SARS-CoV-2 infection in HEC50B cells. (f and g) Effects of drugs on SARS-CoV-2-induced syncytium formation in HEC50B (f) and HEC50B-TMPRSS2 (g) cells. Cells were stained with anti-SARS-CoV-2 N antibody (green) 24 h after infection. Nuclei were stained with Hoechst 33342 (blue). Scale bars, 200 μm. (h and i) Effects of drugs on SARS-CoV-2-induced cytopathicity in HEC50B (h) and HEC50B-TMPRSS2 (i) cells. marima, marimastat; prinoma, prinomastat; E-64d, 25 μM E-64d; nafamo, 10 μM nafamostat. Values are means ± SD (n = 3/group). **, P < 0.01.