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. 2020 Jun 10;10:9369. doi: 10.1038/s41598-020-66055-0

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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 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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Prolonged culture but not LIV decreases proteins associated with oxidoreductase activity. (a) In vitro expansion of MSCs significantly decreased nine proteins related to oxidoreductase activity in LP cells (p < 0.05). G6PD and ME-1 were significantly upregulated in LP + LIV compared to LP (^Ϯp < 0.05) (b) Ten proteins associated with oxidoreductase activity exhibited significantly higher levels in the LP + LIV group compared to LP (p < 0.05) but were not changed compared to P6. Further, xanthine dehydrogenase and ERO1-like protein alpha were significantly elevated in the LP + LIV group when compared to the P6 group (p < 0.05). Identified differences (p < 0.05) between LP and LP + LIV were further denoted as Ϯ: LP + LIV > LP. (c) Representative western blots using primary antibodies against ME-1, G6PD and β-tubulin. Red dashed line indicates a cropped blot, full-length blots are presented in Supplementary Fig.S2. Samples derive from the same experiment and that blots were processed in parallel. (d) All measurements were normalized to P6 (i.e P6 was set to 1). Protein levels in the LP group was 0.63, a 37% decrease compared to the P6 group (p < 0.05). Average ME-1 protein level of the LP + LIV group was 1.36, a 36% (p < 0.05) and a 215% (p < 0.01) increase compared to the P6 and the LP groups. (e) Total G6PD protein level of the LP group was 0.65, remaining 35% lower compared to P6 (p < 0.05). G6PD levels of the LP + LIV group was back to 1.03 and was not significantly different than the P6 group, while remaining 58% larger than the LP group (p < 0.05). n = 3/grp. Group comparisons were made using one-way ANOVA followed by a Newman-Keuls post-hoc test. p < 0.05, **p < 0.01, ***p < 0.001, against control and each other.