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. 2017 Sep 4;7:10393. doi: 10.1038/s41598-017-10927-5

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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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MDM2 binds with NICD1 in GMCs exposed to high glucose and regulates the ubiquination status of NICD1. (A) Cell lysates of high glucose-cultured HBZY-1cells were incubated with MDM2 primary antibody, and NICD1 was pulled down by MDM2 primary antibody without Numb binding. Input: whole lysate; IP: immunoprecipitation; IgG: mouse IgG. (B) Cell lysates of HBZY-1 cells transfected with MDM2 siRNA were incubated with Notch1 primary antibody, and ubiquitination was detected by immunoblotting with anti-ubiquitin antibody. Ub-NICD1 represents ubiquitinated NICD1. (C) Summarized data showing that the ubiquitination NICD1 is regulated by MDM2 inhibition n = 4. ^* P < 0.05 vs. Ctrl + Scra; ^& P < 0.05 vs. HG + Scra.