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. 2022 Aug 25;4(9):1109–1118. doi: 10.1038/s42255-022-00615-8

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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. 3 — a, Left, representative molecular histology of cortical and outer stripe of outer medulla areas of sham kidney (n = 3), generated from three-dimensional UMAP analysis on the basis of lipid profiles recorded by MALDI-MSI (5 × 5 μm² pixel size). Right, LTL immunofluorescent staining on post-MALDI-MSI tissue (red). b, 2D UMAP plot and representative spatial segmentation showing lipid heterogeneity between LTL-positive proximal tubular cells from the cortical and outer stripe of outer medulla areas of sham kidneys (n = 3). c, Representative images showing lipid species distribution in the cortical and outer stripe of outer medulla areas of sham kidney (n = 3), as recorded by MALDI-MSI (5 × 5 μm² pixel size). d, Dynamic metabolic measurements using [U-¹³C₆]glucose on PT cells of sham kidneys. e, Dynamic metabolic measurements using [U-¹³C₅]glutamine on PT cells of sham kidneys. f, Dynamic metabolic measurements using [U-¹³C₁₈]linoleate on PT cells of sham kidneys. Images showing the average ¹³C enrichment of isotopologues on tissue. Graphs showing the comparison of the average ¹³C enrichment of isotopologues between PT S1/S2 and PT S3. The average ¹³C enrichment (area under curve (AUC) normalized to total time) of isotopologues were derived from [U-¹³C₆]glucose or [U-¹³C₅]glutamine measured at different timepoints (0, 15, 30, 60, and 120 minutes) or [U-¹³C₁₈]linoleate measured at different timepoints (0, 60, and 120 minutes). Charts showing the traced isotopes and their derived isotopologues. Two-tailed paired t-test was performed. All scale bars, 200 µm. g, Direct carbon contribution of different nutrients to glutamate at the 2-hour timepoint, as measured from the glutamate isotopologues M+2, M+3, and M+5. One-way analysis of variance (ANOVA) was performed (n = 3). Glc, glucose; 3PG, 3-phosphoglycerate; G3P, glycerol-3-phosphate; R5P/X5P, ribulose-5-phosphate/xylulose-5-phosphate; α-KG, alpha-ketoglutaric acid; Glu, glutamate; Gln, Glutamine; Asp, aspartate.

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