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. 2019 Mar 14;9:4538. doi: 10.1038/s41598-019-40633-3

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© The Author(s) 2019

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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Cold exposure induces PPARα expression through inhibition of the UPS. (a) PPARα and D6D expression under treatment with 20 μM MG132. MG132 was added 6 h after initiation of starvation. (n = 3). (b) FAME analyses of (15 °C) or (37 °C) cells overexpressing ubiquitin. To determine the UPS involvement, cells were treated with 20 μM MG132 for 12 h before collection (n = 3). (c) Ubiquitination of proteins in untransfected cells (Non-ovx), cells overexpressing vector (Vec-ovx), or overexpressing ubiquitin (Ub-ovx). All cells were treated with 20 μM MG132 at 12 h before collection (n = 3). (d) PPARα and D6D expression in untransfected cells, cells overexpressing vector (Vec-ovx), and cells overexpressing ubiquitin (Ub-ovx) (n = 3). (e) Influence of ubiquitin overexpression on cell viability in (15 °C) or (37 °C) cells (n = 3). Assays for (c–e) were carried out at 18 h after initiation of starvation. Blots for PPARα, D6D, Actin, and ubiquitin were derived from individual gels, using aliquots of the same samples. Error bars represent SD. *P < 0.05, **P < 0.01 by unpaired two-tailed Student’s t-test. For gel source data, see Supplementary Fig. 4.