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. 2021 Feb 5;102(3):001531. doi: 10.1099/jgv.0.001531

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© 2021 The Authors

This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.

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Fig. 1. — Generation and validation of H9N2 viruses with mutant PA-X proteins. A panel of mutations were made within UDL-01 segment three that altered PA-X expression. (a) Location of mutations within frameshift site and X-ORF. Dark grey rectangle represents PA, light grey rectangle represents X-ORF. Pink line represents location of the frameshift site (FS), coloured lines represents location of PTC mutations. (b) Coupled in vitro transcription-translation reactions radiolabelled with ³⁵S-methionine were carried out using the TnT rabbit reticulocyte lysate system and protein products analysed using a 15 % SDS-PAGE gel and autoradiography. Red asterisks indicate PA-X polypeptides. PA is marked via a black arrow. (c) Quantification of the AUC of the densitometry analysis of the PA band using ImageJ analysis software. (d) Quantification of the AUC of the densitometry analysis of the PA-X band using ImageJ analysis software in the area indicated by the bracket and compared to local background.