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. 2021 Jan 8;11:232. doi: 10.1038/s41598-020-80291-4

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 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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Glut1 expression is induced by LPS in primary macrophages (Bone marrow derived macrophages, BMDM). (A,B) BMDM cells were treated by HOTAIR siRNA and scramble siRNA followed by LPS treatment and RNA was extracted. The expression of HOTAIR and Glut1 was measured by real time PCR. Data represent mean ± SD (n = 3); *p < 0.05, **p < 0.001, ***p < 0.0001. (C,D) BMDM cells were treated by HOTAIR siRNA and scramble siRNA and treated with LPS. Proteins were analyzed by Western blotting using antibodies against phospho-p65 (NF-κB subunit), IκBα, Glut1, and β-actin (loading control). The changes in amounts of NF-κB, IκBα, and Glut1 have been quantified by ImageLab5.2.1 software is shown in panel (D). (E) BMDM cells were treated with HOTAIR and scramble-siRNA for 48 h and after overnight incubation, cells were stimulated with insulin (+/−), followed by 2-deoxyglucose addition for 20 min. 2-DG6P was oxidized and that generates NADPH, which was quantified calorimetrically (measured at 412 nm in a microplate reader). Each experiment was repeated at least thrice (n = 3). Data represent mean ± SD; *p < 0.05, **p < 0.001.