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. 2022 Sep 12;4(9):1150–1165. doi: 10.1038/s42255-022-00629-2

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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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Extended Data Fig. 3 — a, H&E staining was performed to locate islets (light pink areas, arrows) in Nnat⁺^/-p pancreata. Scale bar, 500 µm. The experiment was repeated independently 3 times with similar results. b, Islet area was quantified as percent of the entire pancreas area. Each group had 3 thin sections from at least 6 animals. All data were plotted as mean ± SEM, *** (adjusted p < 0.0001) by one-sided Tukey’s multiple comparisons test. c, Total insulin content was extracted from whole pancreata, normalized to total insulin in WT littermates. Each group had at least 5 animals. All data were plotted as mean ± SEM, *** (adjusted p = 0.0008) by one-sided Tukey’s multiple comparisons test. d, Glucagon and somatostatin staining was performed on Nnat⁺^/-p pancreata. Scale bar, 100 µm. The experiment was repeated independently 3 times with similar results. e, Cell death (apoptosis) event was examined via TUNEL assay in the pancreatic section from Nnat⁺^/-p and WT littermates at 16 weeks old. DNase treated sample as positive control from the same animals. The black circle heights the islet area. Each group had at least 3 animals. Scale bar, 100 μm f, Basal insulin secretion was measured from size-matched β-cell spheroids. Each group had 8 spheroids. All data were plotted as mean ± SEM. g, h Glucose-stimulated insulin secretion assays were performed on primary islets (g) and spheroids (h) from 16 weeks old Nnat⁺^/-p and WT littermates. 2.8 mM glu.: 2.8 mM glucose and 16.7 mM glu.: 16.7 mM glucose. At least 6 primary islets and 4 spheroids were in each group. All data were plotted as mean ± SEM. i, Oral glucose tolerance test (OGTT) was performed in 16 weeks old Nnat⁺^/-p and WT mice (n = 4-5) fasted for 6 hours and showed relatively normal glucose tolerance in Nnat⁺^/-p-Heavy morphs. All data were plotted as mean ± SEM. j, Growth trajectories for untreated and STZ-treated Nnat⁺^/-p animals and WT littermates between 4 and 12 weeks of age. All data were plotted as mean ± SEM.

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