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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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Pull-down assay with the PDX-1 promoter in INS-1 cells treated with glucose. One mg of nuclear extract from INS-1 cells, who had been treated with 0 mM, 5 mM or 25 mM glucose for 4 h, was incubated with 1 μg of a biotinylated PDX-1 DNA probe. The DNA-protein complex was passed through a μMacs™-column and loaded on a 10% SDS polyacrylamide gel for Western blot analysis. (a) Identification of USF-binding at the PDX-1 promoter was performed with the USF1 specific antibody sc-8983 and with the USF2 specific antibody sc-862. PDX-1 was visualized with the polyclonal rabbit antiserum against recombinant full-length mouse PDX-1. Full-length blots are presented in Supplementary Figure S2. (b) Relative protein amounts of USF1, USF2 and PDX-1 bound to the PDX-1 promoter DNA. The diagram shows the mean ± SD of three independent experiments. Statistical analysis was performed by using Students t-test. *Significant difference p < 0.05, **significant difference p < 0.01, ***significant difference p < 0.001.