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. 2021 Jan 12;11:821. doi: 10.1038/s41598-020-80896-9

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Anti-SARS-CoV-2 activity of λ-CGN. (A) SARS-CoV-2-infected Vero cells (MOI, 0.005) were treated with λ-CGN or RDV for 2 days. Whole cell lysates were subjected to immunoblotting with anti-SARS-CoV-2 spike antibody (upper panel), in which β-actin was used as a loading control (lower panel). Proteins are marked on the right side of the panels. No virus, cell lysates without SARS-CoV-2 infection; Mock, SARS-CoV-2-infected cell lysates without antiviral compound treatment. S2, S2 subunit of viral spike protein. (B) Viral RNA was purified from the culture supernatants of the samples as mentioned in (A). Real-time RT-PCR was conducted using SARS-CoV-2 N gene-specific primers. Relative viral RNA copies were calculated on the basis of their Cq values. (C) Culture supernatants used in (B) were serially diluted for infection of fresh Vero cells. On day 2, the cell culture plates were subjected to immunofluorescence assay for determination of infectious viral titer by calculating the relative ratio of spike-derived green fluorescence frequency to nuclei-derived blue-positive cell number. Data are expressed as the mean ± S.D. of three independent experiments. **P < 0.01; ****P < 0.0001; n.d., not detected. The graphs were created using GraphPad Prism 8.3.1 (www.graphpad.com).