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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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PIAA induces proliferation of rat and human β-cells. (A-B’) Confocal images of rat INS-1 β-cells treated with DMSO (A-A’) and PIAA (B-B’), respectively, stained for Ki67 (red) and Insulin (green). (C) The percentage (mean ± SD) of Ki67 and Insulin-double positive cells (in A-B’; 43.1 ± 5.2% (DMSO) and 87.3 ± 10.4% (PIAA)). (D) Quantification of cAMP levels (mean ± SD) (5.8 ± 0.2 pmol/well (DMSO) and 8.7 ± 0.4 pmol/well (PIAA)). (E) representative Western blot showing increased phosphorylation of PKA substrate and ERK1/2 in PIAA-treated INS-1-derived 832/13 β-cells. Note that PIAA-treatment augmented phosphorylation of mTOR targets RPS6 and Grb10 but did not trigger phosphorylation of AKT^T308 and AKT^S473. (F-G’) Confocal single-plane images of whole rat islets treated with DMSO (F-F’) and PIAA (G-G’), respectively, stained for Ki67 (red, white arrows) and Insulin (green). (H) The percentage (mean ± SD) of Ki67 and Insulin-double positive cells in whole rat islets increased in a dose-dependent manner with treatment of PIAA (1.1 ± 0.3% (DMSO), 2.7 ± 0.9% (20 μM), 3.7 ± 1.5% (40 μM), and 5.5 ± 0.6% (80 μM)). n = 5 replicates per condition. (I) Glucose stimulation indices of rat islets treated with DMSO or PIAA (300 islet equivalents per column, triplicate). (J-K’) Confocal single-plane images of human islets treated with DMSO (J-J’) and PIAA (K-K’), respectively, stained for Ki67 (red, white arrows), Topro (blue), and INSULIN (green). (L) The percentage (mean ± SD) of Ki67 and Insulin-double positive cells in human islets increased in a dose-dependent manner with treatment of PIAA (0.1 ± 0.0% (DMSO), 0.3 ± 0.1% (20 μM), 0.4 ± 0.1% (40 μM), and 0.5 ± 0.2% (80 μM)). n = 5 replicates per condition from 3 cadaveric donors. (M) Glucose stimulation indices of human islets treated with DMSO or PIAA (300 islet equivalents per column, triplicate). *P < 0.05; **P < 0.01; ***P < 0.001.