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. 2018 Feb 12;8:2785. doi: 10.1038/s41598-018-20973-2

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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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CD44 depletion increases insulin secretion and insulin content in Min6 cells. (A) Immunofluorescence staining for CD44v and insulin in Min6 cells. The fluorescence (blue) of nuclei counterstained with DAPI is shown in the merged image. Scale bars, 50 µm. (B) RT and real-time PCR analysis of CD44 gene transcripts in Min6 cells transfected with control or CD44 (#1 or #2) siRNAs. (C) Immunoblot analysis of CD44v and β-actin (loading control) in Min6 cells transfected as in (B). (D) Min6 cells transfected as in (B) were incubated for 1 h in the presence of 5 or 25 mM glucose or of 5 mM glucose (Glc) plus 30 mM KCl, after which the amount of insulin released into the incubation medium was determined with an ELISA. (E) The insulin content of Min6 cells transfected as in (B) was determined with an ELISA before and after incubation for 1 h in the presence of 5 or 25 mM glucose. All quantitative data are means ± s.d. for three independent experiments. *P < 0.05, **P < 0.01 (unpaired Student’s t test).