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. 2019 Jun 6;10:2474. doi: 10.1038/s41467-019-10189-x

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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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Fig. 7 — Hyperglycaemia alters oxidative metabolism in INS-1 832/13 cells. a Abundance of the indicated proteins, quantified by mass spectrometry, in INS-1 cells cultured at 5 mM (black, Ctrl) or 25 mM (white, highG) glucose. Mean ± s.e.m. (n = 4 replicates) and individual data points are indicated. *p < 0.05, **p < 0.01, ***p < 0.001. Student’s t test (unpaired, two-sided). Above, enzymes involved in glycolysis. GPI, glucose 6-phosphate isomerase. PFKL, 6-phosphofructokinase, liver type. ALDOA, aldolase A. GAPDH, Glyceraldehyde 3-phosphate dehydrogenase. ENO1, enolase 1. ENO3, enolase 3. Below, mitochondrial enzymes. DLAT, dihydrolipoyllysine-residue acetyltransferase (a component of the pyruvate dehydrogenase complex). CS, citrate synthase. ACO2, aconitase 2. NDUFA9 and NDUF13, subunits 9 and 13 of NADH dehydrogenase:ubiquinone oxidoreductase (Complex 1). COX6B1, Cytochrome c oxidase subunit 6B1. b, c Oxygen consumption rate (OCR) in INS-1 cells cultured for 48 h at 5 mM (black) or 25 mM (red) glucose, expressed as absolute values (b) and as the percentage of change from the OCR baseline in 2 mM glucose (c). Data are mean ± s.e.m., n = 10 replicates/group (2 and 20 mM Glucose); n = 5, other compounds. d Percentage of change in OCR when glucose was raised from 2 to 20 mM (20G, n = 10 replicates/group), ATP-linked OCR (oligo, n = 5 replicates/group), OCR required to maintain the mitochondrial leak (rot + ant, n = 5 replicates/group) and non-mitochondrial OCR (non-mito, n = 5 replicates/group) in INS-1 832/13 cells cultured for 48 h at 5 mM glucose (hatched, lowG) or 25 mM glucose (white, highG). 20 G, 20 mM glucose. Oligo, 5 µM oligomycine. Rot + Ant, 50 µM rotenone + 5 µM antimycin A. Mean ± s.e.m. Student’s t test (unpaired, two-sided). *p < 0.05; **p < 0.01; ***p < 0.001. e Insulin secretion, expressed as a percentage of insulin content, for INS-1 cells cultured for 48 h at 5 mM (lowG, hatched) or 25 mM (highG, white) glucose and then exposed to 2 or 20 mM glucose. Mean ± s.e.m. (n = 6 experiments). Two-way analysis of variance **p < 0.01