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. 2020 Aug 5;5:73. doi: 10.1038/s41541-020-00220-4

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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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Fig. 3 — a IFA analysis of JEV-ΔNS1 virus at different passages using 4G2 monoclonal antibody. The JEV-ΔNS1 virus generated through transfection of BHK_NS1 cells with the transcribed JEV-ΔNS1 genomic RNA was designated as P0 virus. Three JEV-ΔNS1 virus stocks (A, B and C) were blind passaged independently for 15 rounds (P0−P15) in BHK_NS1 cells. The viruses at P0 and P15 were used to infect BHK_NS1 cells and naïve BHK-21 cells. JEV-WT was used as a positive control. The length of the scale bar (displayed in a red line segment) represents 20 μm. b RT-PCR analysis of the expression of NS1 gene in BHK_NS1 cells infected with P0 or P15 JEV-ΔNS1 viruses. The uncropped and unprocessed gel including markers was displayed in Supplementary Fig. 2. c Sequence chromatograms of RT-PCR products containing NS1 fragment amplified from P15 JEV-ΔNS1 virus.