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. 2021 Feb 24;11:4494. doi: 10.1038/s41598-020-80425-8

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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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Protein expression levels showed a trend similar to those of the mRNA levels. (a) Normal mice were sacrificed after SHH5000 or SHH8000 treatment for 16 h. Each cerebellum was collected and immediately stored in liquid nitrogen. Western blotting was then performed to detect differences in mitochondrial-related protein expression levels (Mfn2, Opa1, TFAM, UCP4). (b) Western blotting was performed to detect differences in mitochondrial-related protein expression levels (Mfn2, Opa1, TFAM) in each mouse hippocampus after SHH5000 or SHH8000 treatment. (c) Western blotting was performed to detect differences in mitochondrial-related protein expression levels (Mfn2, Opa1, TFAM) in the muscles of mice after SHH5000 or SHH8000 treatment. Data are analyzed by Image J software, and are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the control group, n = 5 per group. Full-length blots/gels are presented in Supplementary Fig. 1a–c. Mfn2, mitochondrial fusion protein 2, a mitochondrial fusion factor on the mitochondrial outer membrane; Opa1, Optic atrophy1, a mitochondrial fusion factor on the mitochondrial inner membrane; TFAM, mitochondrial transcription factor; UCP4, uncoupling protein 4, a mitochondrial stress factor; SHH5000,simulated hypobaric hypoxia at an altitude of 5000 m; SHH8000, simulated hypobaric hypoxia at an altitude of 8000 m.