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. 2022 Mar 3;40(7):1042–1055. doi: 10.1038/s41587-022-01219-z

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© The Author(s) 2022

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. 4 — a, Left, Overview of experimental setup. SC-islets or adult islets were exposed to low (3 mM) or high (17 mM) concentrations of uniformly labeled [U-¹³C6] glucose for 1 h before metabolite extraction and liquid-chromatography mass spectrometry (LC-MS) detection. Right, An example of isotopologue nomenclature and glucose-derived labeling of downstream metabolites. b, The ratio of M+6 G6P to M+6 labeled glucose under low and high glucose concentrations in adult islets and SC-islets over 6 weeks of maturation. c, The relative abundances of fully labeled glycolytic intermediates in SC-islets and adult islets following low and high labeled glucose treatment. d, The M+3 lactate content of adult islets and SC-islets detected over the timecourse of maturation, following low and high labeled glucose treatment. e, The percentage of total serine and glycine labeled from ¹³C-glucose following low and high glucose treatment of SC- and adult islets. f, Ratiometric analysis of labeled lactate to labeled pyruvate, and labeled citrate to labeled pyruvate under high glucose treatment in SC-islets and adult islets. g, The combined percentage of labeled TCA metabolites (M+2 to fully labeled M+n) from SC- and adult islets after low and high labeled glucose treatment. h, The combined abundance of aspartate (M+2 to M+4) and glutamate (M+2 to M+5) isotopologues in SC-islets (w0–w6) under low and high glucose concentrations, relative to adult islets. i, The combined relative abundance of M+2 to M+5 GSH isotopologues under low and high labeled glucose concentrations in adult islets and SC-islets. j, Schematic overview of active glucose metabolic pathways in SC-islets and adult islets. Arrow thickness denotes the extent of glucose-derived carbons entering the pathway. Error bars ± s.e.m. with statistical significance determined by two-tailed t-tests. Hash symbols indicate internal significance from low to high glucose labeling, asterisks denote significance between SC-islet timepoints or adult islet samples at each glucose concentration. ^#,*P < 0.05, ^##,**P < 0.01, ^###,***P < 0.001. SC-islets S7w0 (n = 4), S7w3 (n = 12–13), S7w6 (n = 3), adult islets (n = 6).