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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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Characterization of JEV, YFV and CHKV protein expression and VLP release. (A) 293 T cells were transfected with JEV CprME expression vector. Cells were analyzed for E protein and Capsid protein expression after staining with respective antibodies followed by fluorescence microscopy. (B) 293 T cells were transfected with JEV CprME expression vector alone or the JEV CprME vector along with Zika NS2B-3 expression plasmid. Culture supernatants were ultracentrifuged and analyzed for E protein and Capsid protein expression by western blotting. (C) 293 T cells were transfected with YFV CprME expression vector and analyzed for E protein and Capsid protein expression by immunofluorescence microscopy. (D) 293 T cells were transfected with YFV CprME expression vector alone or the YFV CprME vector along with Zika NS2B-3 expression plasmid. Culture supernatants were ultracentrifuged and analyzed for E protein and Capsid protein expression by western blotting. (E) 293 T cells were transfected with CHKV expression vector and analyzed for E1-E2 protein expression by immunofluorescence microscopy. (F) 293 T cells were transfected with CHKV expression vector. Culture supernatants were ultracentrifuged and analyzed for E1-E2 protein expression by western blotting. Images A, C and E were analyzed using NIS Elements AR software version 3.2 (Nikon; https://nis-elements-viewer.software.informer.com/3.2/). Images B, D and F were analyzed using GENETOOLS gel analysis Software version 4.03 (f) (Syngene, https://www.syngene.com/software/genetools-automatic-image-analysis/).