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. 2017 Nov 27;7:16367. doi: 10.1038/s41598-017-16590-0

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© The Author(s) 2017

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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Influence of glucose on the transcription of the PDX-1 promoter of INS-1 cells. (a) INS-1 cells were transfected with the PDX-1 promoter construct −6500/+68-STF-luc and starved overnight. The next day, cells were treated with 0 mM, 5 mM, 11 mM or 25 mM glucose and harvested after a period of 4 h. Luciferase activity was determined in triplicate; the activity in the cells treated with 0 mM glucose was set to 100%. Statistical analysis was performed by using Students t-test. *Significant difference p < 0.05. (b) INS-1 cells were starved overnight and the next day treated with 0 mM, 5 mM, 11 mM or 25 mM glucose and harvested after a period of 24 h for mRNA isolation with TRIzol® Reagent. PDX-1 mRNA levels were detected semi-quantitatively by real-time RT-PCR. Fold change of PDX-1 mRNA expression relative to 0 mM glucose is displayed (mean ± SD, n = 3). (c) INS-1 cells were transfected with the PDX-1 promoter construct −6500/+68-STF-luc and the USF1 expression plasmid. After overnight starvation, cells were treated with 0 mM, 5 mM, 11 mM or 25 mM glucose and harvested after a period of 4 hours. Luciferase activity was determined in triplicate; the activity in the 0 mM glucose treated cells was set to 100%. Statistical analysis was performed by using Students t-test. *Significant difference p < 0.05. (d) The corresponding Western blot analysis of the FLAG-tagged USF1 is shown beside the graph. Identification of FLAG-tagged USF1 was performed with the mouse monoclonal antibody FLAG M2 (F1804), and α-tubulin served as a loading control. Full-length blots are presented in Supplementary Figure S2.