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. 2018 Oct 22;8:15587. doi: 10.1038/s41598-018-33875-0

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© The Author(s) 2018

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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TBK1/IKKε inhibitors have modest effects on α-to-β-cell transdifferentiation but strongly enhance β-cell proliferation. (A–C) Confocal images of [Tg(ins:CFP-NTR)^s892; Tg(ins:Kaede)^jh6] larvae at 24 hpa, concurrently treated with EdU and DMSO (A), amlexanox (B), or PIAA (C), respectively, from 0–24 hpa, stained for Somatostatin (blue). (D) Quantification of the number (mean ± SD) of Insulin and Somatostatin-double positive cells at 24 hpa (in A-C; 0.7 ± 0.6 (DMSO), 1.7 ± 0.8 (amlexanox), and 1.8 ± 0.5 (PIAA)). (E–G) Confocal images of [Tg(ins:CFP-NTR)^s892; Tg(ins:Kaede)^jh6] larvae at 24 hpa, concurrently treated with EdU and DMSO (E), amlexanox (F), or PIAA (G), respectively, from 0–24 hpa, stained for Glucagon (blue). Note that the number of Insulin and Glucagon-double positive cells (blue arrows) was increased in TBK1/IKKε-I-treated recovering larvae (F, G) compared to DMSO-treated larvae (E). PIAA-treated larvae also showed an EdU-incorporated β-cell (white arrow) (G). (H) Quantification of the number (mean ± SD) of Insulin and Glucagon-double positive cells at 24 hpa (in E-G; 1.5 ± 1.1 (DMSO), 3.4 ± 1.3 (amlexanox), and 4.4 ± 1.9 (PIAA)). (I–K) Confocal images of [Tg(ins:CFP-NTR)^s892; Tg(ins:Kaede)^jh6] larvae at 48 hpa, concurrently treated with EdU and DMSO (I), amlexanox (J), or PIAA (K), respectively, from 24–48 hpa. The number of β-cells that incorporated EdU (white arrows) was significantly increased in TBK1/IKKε-I-treated recovering larvae (J, K) compared to DMSO-treated larvae (I). (L) The percentage (mean ± SD) of regenerated β-cells that incorporated EdU at 48 hpa (in I-K; 4.0 ± 7.0% (DMSO), 32.0 ± 6.0% (amlexanox), and 39.0 ± 6.0% (PIAA)). Cells in 20 planes of confocal images from 10 individual larvae were counted per condition. **P < 0.01.