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. 2020 Nov 24;10:20482. doi: 10.1038/s41598-020-77621-x

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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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Silencing the expression of DNAB3 abolishes the protective effect of ALA on tunicamycin-induced ER stress in C2C12 cells. (A,B) Knocking down the expression of DNAJB3 and HSP72 with 20 nM of specific siRNA blunted the endogenous expression of DNAJB3 and HSP72 mRNAs (A) and proteins (B). Actin was used as internal control for both RT-PCR and western blots. Full-length blots are displayed in Supplementary Fig. S4. ALA fails to protect siRNA DNAJB3-transfected C2C12 cells from tunicamycin-induced mRNA expression of ER stress markers (C) and ATF6-dependent luciferase activity (D). Ethanol and DMSO were used at 0.25% as vehicles for ALA and tunicamycin, respectively. After performing Shapiro–Wilk normality test, t test was used to compare the difference between ALA and vehicle (A) and two-ways ANOVA was used to compare the effect of treatments (B,C). NS not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ^#P < 0.0001.