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. 2018 Feb 2;9:485. doi: 10.1038/s41467-018-02939-0

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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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Fig. 1 — c-Myc plays a role in β-cell proliferation and function. a Cell density of INS-1 cells upon Myc inhibition with 40 µM Myci for 2 days as compared to control (DMSO) treatment. Scale bar, 100 μm. b Quantitative PCR to detect gene expression of key β-cell regulatory transcription factors upon Myc inhibition (Myci, 40 µM) after 2 days of inhibitor treatment, n = 3 per group. **p < 0.005, ***p < 0.0005, Student’s t test. c Control (DMSO, n = 4) or Myci-treated (40 µM, 3 days, n = 4) INS-1 cells were subjected to glucose-stimulated insulin secretion (GSIS) under basal (2.8 mM glucose) followed by stimulatory (16.7 mM with 100 µM IBMX) conditions. *p < 0.05, **p < 0.005, ***p < 0.0005, Student’s t test. d Fold change in GSIS in INS-1 cells treated with Myci (n = 4) as compared to controls (n = 4). Error bars indicate ± SD. **p < 0.005, Student’s t test. e Cell density of INS-1 cells transfected with siMyc as compared to control samples transfected with a scrambled siRNA (siScr) for 5 days. Scale bar, 100 μm. f Quantitative PCR analysis of INS-1 cells to evaluate gene expression of several β-cell transcription factors upon reduction of c-Myc (siScr, n = 6, siMyc, n = 4–6). *p < 0.05, ***p < 0.0005, Student’s t test. g Secretory response in INS-1 cells depleted of c-Myc (siMyc) as compared to control (siScr) cells. n = 3 per group. **p < 0.005, Student’s t test. h GSIS measured in INS-1 cells with siMyc as compared to siScr samples. n = 3 per group. Error bars indicate ± SD, p = 0.05, Student’s t test. i Western blot analysis of Myc levels in juvenile (3 weeks old, n = 5) islets versus adult (3 months old, n = 6) islets from wild-type mice. p = 0.08, Student’s t test. j Quantitative PCR of β-cell maturation genes in adult islets (3 months old, n = 5) as compared to juvenile (3-weeks-old islets, n = 5) along with a cell cycle gene. *p < 0.05, **p < 0.005, Student’s t test. k BrdU incorporation (expressed as %BrdU per Insulin + ve cells) in p16 pups to quantify actively replicating β cells in the transgenic (Ins-Cre;Myc^+/−, n = 3 or Ins-Cre;Myc^−/−, n = 4) animals as compared to control (n = 4) littermates. ***p < 0.0005, Student’s t test