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. 2020 Dec 4;10:21226. doi: 10.1038/s41598-020-78210-8

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© The Author(s) 2020

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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HA proteolytic activation and replication of recombinant viruses in different MDCK cell lines. Cleavage-activation of the HA of recombinant viruses carrying single mutations in the HACS in CEK cells with ( +) or without ( +) trypsin (A), in CEK after transfection with TMPRSS2 ( +) or left untransfected (-) (B) or MDCK cells expressing HAT (C). Western Blot figures were acquired by Quantity One Software Version 4.4 (Biorad, Germany) (https://www.bio-rad.com/webroot/web/pdf/lsr/literature/4000126-14A.pdf). Multiple cycle replication after infection of MDCK with or without trypsin, MDCK-TMPRSS2 or MDCK-HAT with different recombinant viruses at MOI of 1 for 24 h. Titers were determined by plaque assay in MDCKII cells in the presence of trypsin. Titers are expressed as PFU/ml and are shown as the mean and the standard deviation. The kinetics were done twice in duplicates for each cell types (D). Data were analyzed using one-way ANOVA with post hoc Tukey tests and Kruskal–Wallis with Dunn’s test. Asterisk indicates significant differences compared to K319 (p < 0.05).