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. 2020 Mar 4;10:4017. doi: 10.1038/s41598-020-61103-1

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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 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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Generation of stable cell lines secreting JEV, YFV and CHIKV VLPs. 293 T cells were transduced with (A) JEV (B) YFV or (C) CHKV structural protein containing lentivirus particles. Transduced cells were bulk selected with blasticidin followed by limiting dilution single cell cloning. For each virus, several single cell clones were characterized for Envelope protein staining on the cell surface via flow cytometry and VLP secretion into the culture supernatants via western blotting. Mean Fluorescence intensity (MFI) of Envelope protein staining for the different single cell clones for JEV, YFV and CHIKV is shown on the right. Gel images were analyzed using GENETOOLS gel analysis Software version 4.03 (f) (Syngene, https://www.syngene.com/software/genetools-automatic-image-analysis/). Flow cytometry histograms were created using the FloJo Software version 10.6.0 (Tree Star, https://www.flowjo.com/solutions/flowjo/downloads).