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. 2020 Nov 4;10:19068. doi: 10.1038/s41598-020-76227-7

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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CVB3-induced LC3 lipidation is independent of BECN1 and PIK3C3. (A) BECN1-KO HEK293A cells were established through CRISPR-cas9 editing. Knockout efficiency was verified by western blotting (left panel). WT or BECN1-KO cells were cultured in either normal medium, HBSS starvation medium, or starvation medium supplemented with 200 nM BAF for 2 h, followed by western blot analysis of LC3 (middle panel). WT or BECN1-KO cells were sham- or CVB3-infected for 16 h, followed by western blot assessment of LC3 (right panel). (B,C) BECN1 (B) and PIK3C3 (C) were transiently silenced in HEK293A cells via siRNA treatment for 48 h. Cells were then subjected to sham or CVB3 infection for 16 h. Cells were harvested and subjected to western blot analysis of LC3, BECN1, and PIK3C3. Densitometric results are presented either underneath the blots or in the right panel (mean ± SD, n = 3, analyzed by one-way ANOVA with Tukey’s post-test).