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. 2025 Apr 9;44(10):2856–2881. doi: 10.1038/s44318-025-00434-z

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© The Author(s) 2025

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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) Schematic diagram of the lineage tracing strategy by Ngn3-CreER;R26-tdT mice. (B) Schematic showing the experimental strategy. (C) Wholemount fluorescent images of pancreas collected from Ngn3-CreER;R26-tdT at the indicated time points after Tam treatment. Arrowheads, tdT⁺ pancreatic islets. (D) Immunostaining for tdT and CK19 or Ins or somatostatin (Sst) on pancreatic sections of Ngn3-CreER;R26-tdT. (E) Immunostaining for tdT and Ins or Sst or glucagon (Gcg) or pancreatic polypeptide (Ppy) on pancreatic sections of Ngn3-CreER;R26-tdT mice. Arrowheads, tdT⁺lineage⁺ pancreatic endocrine cells. (F) Quantification of the percentage of tdT⁺ cells among pancreatic endocrine cell lineages in the Ngn3-CreER;R26-tdT mice. Data are mean ± SD; +2 weeks, n = 5 biological replicates; +12 weeks, n = 4 biological replicates; Ins: P = 0.30; Sst: P = 0.72; Gcg: P = 0.97; PP: P = 0.62; n.s., non-significant. In each sample, islets from 10 pancreas sections were quantified. Two-tailed unpaired Student’s t-tests were used for statistical comparisons and P < 0.05 was accepted as statistically significant. (G) Schematic of the Ngn3-2A-CreER knock-in strategy. (H) Schematic of lineage tracing strategy for Ngn3-2A-CreER;R26-tdT. (I) Schematic showing the experimental strategy. (J) Wholemount fluorescent images of the pancreas from Ngn3-2A-CreER;R26-tdT mice at the indicated time-points after Tam treatment. Arrowheads, tdT⁺ pancreatic islets. (K, L) Immunostaining for tdT and E-cad (K) or CK19 or Ins or Sst (L) on pancreatic sections of Ngn3-2A-CreER;R26-tdT mice. Arrowheads in (K), tdT⁺ islets. (M) Immunostaining for tdT and Ins or Sst or Gcg or Ppy on pancreatic sections of Ngn3-2A-CreER;R26-tdT Mice. Arrowheads, tdT⁺lineage⁺ pancreatic endocrine cells. (N) Quantification of the percentage of tdT⁺ cells in pancreatic endocrine cell lineages of Ngn3-2A-CreER;R26-tdT mice. Data are mean ± SD; +2 weeks, n = 5 biological replicates; +12 weeks, n = 5 biological replicates; Ins: P = 0.76; Sst: P = 0.26; Gcg: P = 0.42; PP: P = 0.67; n.s., non-significant. In each sample, islets from 10 pancreas sections were quantified. Two-tailed unpaired Student’s t tests were used for statistical comparisons and P < 0.05 was accepted as statistically significant. (O) Cartoon image showing that Ngn3-CreER and Ngn3-2A-CreER label a subset of ductal cells and endocrine cells in the adult pancreas. Scale bars, 1 mm (yellow) and 100 μm (white). Each image is representative of 4–5 individual mouse samples. See also Fig. EV1. Source data are available online for this figure.