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. 2020 Mar 31;51:48. doi: 10.1186/s13567-020-00771-3

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© The Author(s) 2020

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Assessing the cleavability of the hemagglutinin cleavage site (HACS) of different avian influenza viruses. A A secretable luciferase protein construct was created by linking LUCIA^® luciferase to a trans-Golgi retention anchor (BACE) via an influenzaviral HACS. B Luciferase activity in the supernatant of transfected cell cultures is proportional to cleavage at the HACS within the constructs. The constructs were expressed in avian (QM9, DF-1) and mammalian cells (A549, ST). LoVo cells do not express furin activity [51]. Expression levels are compared quantitatively, error bars represent 1 SD from ten independent experiments; *(p < 0.05; two-sample t-test) indicates a statistically significant difference from the NC-BACE negative control and from other samples. PC_H5_poly represents the polybasic HACS of a highly pathogenic avian influenza virus H5N1 from Bangladesh and is used here as a positive control.