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. 2018 Aug 22;8:12590. doi: 10.1038/s41598-018-30820-z

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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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An in vitro IR model was successfully established in cultured 3T3-L1 adipocytes. (A) Morphological observations and Oil red O staining of 3T3-L1 adipocytes. (B) Glucose consumption test showed low levels of glucose under insulin stimulation in IR cells. (C) Glucose uptake assay showed low levels of glucose uptake under insulin stimulation in IR cells. (D) Immunofluorescence detection revealed inhibition of GluT4 membrane translocation in IR cells. (E) GluT4 expression in the cell membranes decreased in IR cells according to western blot. (F) The semi-quantification of GluT4 (E) by western blot and using ImageJ software. (G) The mRNA of Twist 1 was determined in IR cells based on real-time PCR. (H) The expression of Twist 1 was determined in IR cells based on western blot. (I) The siRNA induced an obvious Twist 1 silencing effect according to western blot. (J) Glucose uptake tests revealed elevated glucose in Twist 1-silenced cells. (K) Glucose consumption assay also showed elevated glucose in Twist 1-silenced cells.