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. 2018 Jun 5;8:8585. doi: 10.1038/s41598-018-26975-4

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© The Author(s) 2018

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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HEV ORF3 impairs endogenous type I interferon production in host cells. Adenovirus and plasmid vectors expressing ORF3 protein were transfected into THP1 and LO2 cell lines, respectively. (A) ELISA data showing that IFNα and IFNβ levels in THP1 cells expressing ORF3 protein are significantly decreased (P_α = 0.0133, P_β = 0.042). (B) mRNA levels of type I interferon were inhibited in ORF3-expressing cells (P_α = 0.0272, P_β = 0.0251). (C) The TLR4 level was not changed while TLR3 and TLR7 levels were significantly reduced in THP1 cells infected with Ad-ORF3 (P_TLR3 = 0.0087, P_TLR4 = 0.1185, P_TLR7 = 0.0038). (D) Expression of TLR3, TLR4 and TLR7 was confirmed in LO2 cells transfected with pcDNA3.1-GFP-ORF3 (P_TLR3 = 0.0285, P_TLR4 = 0.3297, P_TLR7 = 0.0054). (E) Protein levels of TLR3, TLR4 and TLR7 in LO2 cells were detected via western blot. Lane 1: Normal control LO2 cells. Lane 2: LO2 cells transfected with pcDNA3.1-GFP. Lane 3: LO2 cells transfected with pcDNA3.1-GFP-ORF3. Results are presented as means ± SD. Error bars indicate SD. *P < 0.05, **P < 0.01. All of full-length gels and blots are included in Supplement S3.