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. 2020 Apr 22;10:6846. doi: 10.1038/s41598-020-63761-7

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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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Exposure of preadipocytes to chronic intermittent hypoxia (IH) attenuates lipid accumulation during differentiation. Representative images of Oil-Red-O stained preadipocytes differentiated for 6 days (a). Quantitation of lipid content accumulated in differentiated cells (b) (n = 4 independent experiments). Compared to preadipocytes grown in continuous normoxia (NO), preadipocytes exposed to IH show less lipid accumulation at day 6 of differentiation. Representative Western Blots and graphs showing protein expression of peroxisome proliferator-activated receptor γ (PPARγ) (c) (n = 7 independent experiments), cAMP response element-binding (CREB) (d) (n = 8 independent experiments), and hypoxia-inducible factor-1α (HIF-1α) (e) (n = 7 independent experiments) in 6 day differentiated preadipocytes. IH elevated expression of PPARγ and CREB even in absence of exposure to differentiating media. HIF-1α expression was elevated in preadipocytes exposed to IH prior to treatment with differentiating media. 1: NO; 2: NO + differentiation in NO; 3: IH; 4: IH + differentiation in NO; 5: IH + differentiation in IH; 6: etopside + differentiation in NO. Data are presented as mean ± SEM. P-values determined by one-tailed paired t-test compared to the cells grown and differentiated in continuous NO (2). *P < 0.05. Etopside was used as a positive control. The same blot was used for the representative images of PPARγ and CREB. Therefore, the image for GAPDH protein is also the same.