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. 2023 May 10;14:2698. doi: 10.1038/s41467-023-38377-w

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© The Author(s) 2023

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Wild-type and Ube2g2^−/− cells were infected with either R18 labelled ZIKV at MOI 400 or pre-labelled control medium and visualised by immunofluorescence imaging. Hoechst staining indicates nuclei; red puncta structures indicate R18 labelled virus. b The R18 positive population was measured using flow cytometry at 2.5 hpi. Error bars represent mean ± SD, n = 3 biologically independent samples. c Schematic representation of Zika virus replicon; the structural gene segments are substituted with a luciferase reporter for activity readout. d–f Luminescence of ZIKV replicon RNA expressing luciferase in wild-type and Ube2g2^−/− cells measured at indicated time points after transfection to measure translation (d) and virus replication in wild-type, Ube2g2^−/−, Ube2g2^WT and Ube2g2^C89K cells (e, f). Data are presented as mean ± SD, n = 4 biologically independent samples (e, f). Luciferase activity is shown as relative luminescence units (RLU). **P < 0.01; ***P < 0.001 (compared with wild-type by ANOVA followed by one-sided Dunnett’s test). g, h Deletion of Ube2g2 affects the structure of intracellular viral replication and assembly complex. Wild-type and Ube2g2^−/− cells were infected with Zika at MOI of 5. Cells were fixed at 24hpi and probed with anti-ZIKV envelope E-protein (488) and anti-double stranded (ds) RNA (555). Nuclei were visualised by Hoechst staining (g); Scale bar = 10 µM. Pearson’s correlation coefficient was measured to determine co-localisation (h). Error bars represent mean ± SD, n = 6 biologically independent samples; **P < 0.01; ***P < 0.001 (compared with wild-type by ANOVA followed by one-sided Dunnett’s test). (Source data are provided as a Source data file).