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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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Enterovirus A71 induces autophagosome formation in neuronal cells and differentiated IMR-32 cells. Neuronal cells derived from human NSCs and IMR-32 cells were infected with EV-A71 at an MOI of 2. (A) Immunofluorescence images of the LC3 and EV-A71 3D^pol in EV-A71-infected neurons at 12 and 24 h postinfection. The green color is LC3, the red color is EV-A71 3D^pol, and DAPI was used to label nuclei. The scale bar represents 20 μm. (B) The average number of LC3⁺ puncta per cell was quantified according to the immunofluorescence results. (C) Western blot assays were performed to measure the protein levels of LC3-I, LC3-II and EV-A71 3D^pol in EV-A71-infected neurons. (D) The band intensities of LC3-II were measured using ImageJ software and normalized to the β-actin. The time point of 0.5 h was used as a reference. (E) The protein levels of LC3-I, LC3-II and EV-A71 3D^pol of EV-A71-infected differentiated IMR-32 were detected using western blot. (F) The band intensities of LC3-II were measured using ImageJ software and normalized to the β-actin. (G) The protein levels of phospho-Beclin1 S15, total Beclin1 and EV-A71 3D^pol of EV-A71-infected differentiated IMR-32 were detected using western blot. β-actin was used as an internal control in western blots. The error bars represent the SD. Student’s t test. **p < 0.01; ***p < 0.001.