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. 2020 Sep 17;10:15234. doi: 10.1038/s41598-020-71970-3

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© The Author(s) 2020

Open AccessThis 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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The accumulation of autophagosomes increases Enterovirus A71 vRNA replication. (A) Differentiated IMR-32 cells infected with EV-A71 (MOI = 2) and treated with 5 or 10 nM bafilomycin A1 or with 10 μM pepstatin A and E-64d or with DMSO only. Immunofluorescence assay was performed to detect the expression of LC3-positive puncta at 24 h postinfection. Green color is LC3 and DAPI was used to label nuclei. The scale bar represents 10 μm. (B) The average number of LC3⁺ puncta per cell was quantified according to the immunofluorescence results. (C) Differentiated IMR-32 cells infected with EV-A71 (MOI = 2) or went mock-infection, and treated with 5 or 10 nM bafilomycin A1 or with DMSO only. Total RNA was harvested at 12 and 24 h postinfection, and RT-qPCR assays were performed to measure the relative levels of EV-A71 5′ UTR vRNA. (D) Differentiated IMR-32 cells infected with EV-A71 (MOI = 2) or went mock-infection, and treated with 10 μM pepstatin A and E-64d or with DMSO only. Total RNA was harvested at 12 and 24 h postinfection, and RT-qPCR assays were performed to measure the relative levels of EV-A71 5′ UTR vRNA. The experiments were performed in triplicate, and the error bars represent the SD. Student’s t test. *p < 0.05; **p < 0.01; ***p < 0.001.