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. 2024 Dec 19;9:246. doi: 10.1038/s41541-024-01033-5

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 1 — a Schematic representation of the plasmids that encode the segments of the influenza virus used in reverse genetics and the sequence of the plasmid pPR169-PspA-178, which encodes part of the neuraminidase segment together with the PspA4Pro protein. b Hypothetical representation of the different viruses used in the study: wild-type influenza virus A/PR8/34 (PR8), Flu-CT (a recombinant influenza virus that carries a non-coding spacer sequence) and Flu-PspA (a recombinant influenza virus carrying the PspA4Pro protein from S. pneumoniae). c Phenotype of lysis plaques observed in MDCK cells after titration under agarose of A/PR8/34 and the recombinant viruses Flu-CT and Flu-PspA. d In vitro viral multiplication kinetics of recombinant influenza viruses in MDCK cells up to 72 h after infection. e Viral titers obtained with the multiplication of the Flu-PspA recombinant virus in embryonated chicken eggs for 48 h in different concentrations (300 µU or 600 µU) of exogenous neuraminidase. The bars represent the means ± standard deviation of the data, and ANOVA determined differences between groups. **** indicates a statistically significant difference at p value <0.0001, represented above the bars or connecting lines. f Labeling of PspA4Pro protein (in green/FITC) in uninfected MDCK cells (Mock) or infected with the Flu-CT or Flu-PspA recombinant viruses evaluated by confocal microscopy at 20 h after infection.