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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 illustrating the labeling strategy by Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice. (B) Schematic showing the experimental strategy. (C) Wholemount fluorescence images of the pancreas from Ins2-DreER;Hnf1b-2A-CreER;R26-TLR 2 weeks (left) and 12 weeks (right) after Tam treatment. Arrowheads, pancreatic islets. (D) Immunostaining for ZsGreen, tdT, and CK19 on pancreatic sections of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice 2 weeks after Tam treatment. (E) Quantification of the percentage of tdT⁺CK19⁺ cells among CK19⁺ pancreatic ductal cells of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice. Data are mean ± SD; n = 5 biological replicates. In each sample, islets from 10 pancreas sections were quantified. (F) Immunostaining for ZsGreen, tdT, and CK19 or Ins or Sst on pancreatic sections of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice 2 weeks after Tam treatment (G) Immunostaining for ZsGreen, tdT, and Ins on pancreatic sections of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice 2 weeks after Tam treatment. Arrowheads, ZsGreen⁺tdT⁺Ins⁺ pancreatic beta cells. (H) Immunostaining for ZsGreen, tdT, and CK19 or Ins or Sst on pancreatic sections of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice 12 weeks after Tam treatment. (I) Immunostaining for ZsGreen, tdT, and Ins on pancreatic sections of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice 12 weeks after Tam treatment. Arrowheads, ZsGreen⁺tdT⁺Ins⁺ pancreatic beta cells. (J) Quantification of the percentages of ZsGreen^–tdT⁺ and ZsGreen⁺tdT⁺ or ZsGreen⁺ cells in Ins⁺ pancreatic beta cells of Ins2-DreER;Hnf1b-2A-CreER;R26-TLR mice. Data are mean ± SD; +2 weeks, n = 5 biological replicates; +12 weeks, n = 5 biological replicates; ZsGreen⁺tdT⁺: P = 0.27; ZsGreen⁺: P = 0.57; 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. (K) Cartoon image showing that Ins2-DreER;Hnf1b-2A-CreER;R26-TLR label Hnf1b⁺Ins^– non-beta cells, Hnf1b⁺Ins⁺ beta cells, and Hnf1b^–Ins⁺ beta cells in pancreas simultaneously. Hnf1b⁺Ins^– non-beta cells do not contribute to beta cells was detected during homeostasis. Scale bars, 1 mm (yellow) and 100 μm (white). Each image is representative of five individual mouse samples. See also Fig. EV5. Source data are available online for this figure.