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. 2018 Sep 7;8:13418. doi: 10.1038/s41598-018-31734-6

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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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USP4 participates in antiviral immunity against EV71 infection. (A) Analysis of phase contrast microscopy of RD cells transfected with either an empty vector or USP4, and then infected with EV71 (MOI = 0.5) at the indicated time (magnification 20x). (B) Apoptosis assay of EV71-infected RD cells transfected with empty vector or USP4. The apoptosis assay was conducted using annexin V/FITC and PI double staining and flow cytometry. Q3, Q4, and Q2 represent normal cells, early apoptotic cells, and late apoptotic/necrotic cells, respectively. (C,F) q-PCR and western blot analysis of EV71 protein expression in RD cells transfected with control vector or HA-USP4 plasmids, followed by treatment with EV71 for variable lengths of time. (E) Viral titre measurement of EV71 propagating in RD cells transiently transfected with control vector or HA-USP4 plasmids. Data in (D,E) are presented as the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001. The independent experiments were performed in triplicate.