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. 2021 Sep 6;11:17705. doi: 10.1038/s41598-021-97162-1

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© The Author(s) 2021

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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Mortalin levels control neuronal survival against stress induced by rotenone and H₂O₂ and mitochondrial accumulation of ROS. Rat embryonic primary cortical neurons were transduced on DIV 3 with lentiviral vectors (LV) to induce over- (+ 100%, LV-Mortalin) or under- (− 50%, LV-shMortalin, clone A) expression of Mortalin. A lentiviral vector expressing a shRNA with a scramble sequence was used as a control (LV-shScr). (A) On DIV 11, neurons were treated or not with 10 nM rotenone for 24 h, or with hydrogen peroxide (H₂O_2, 100 μM) and then processed the day after. Percentages of cells showing pyknosis (revealed by DAPI staining) are presented as means ± SD of 4 independent experiments. (B) On DIV 11, neurons were treated or not with 100 μM NAC or H₂O₂. After 24 h, mitochondrial ROS accumulation was measured using the MitoSOX dye. Values were normalized using fluorometric values obtained for untreated and non-transduced neurons for each experiment. Results are presented as means ± SD of 3 independent experiments. ****p < 10⁻⁴, by 1-way ANOVA with Sidak’s multiple comparison post-hoc test.