Journal: Molecular Metabolism

Article Title: Dysregulation of the Pdx1/Ovol2/Zeb2 axis in dedifferentiated β-cells triggers the induction of genes associated with epithelial–mesenchymal transition in diabetes

doi: 10.1016/j.molmet.2021.101248

Figure Lengend Snippet: Loss of β-cell identity is associated with alteration of islet microenvironment and TGFβ-dependent fibrosis . (A) Western blotting for phospho-Ser423/425 and γ-tubulin in islets from 12w Tg7 mice. (B) mRNA levels for TGFβ target genes in islets from 12w Tg7 and Wt mice obtained by qPCR (n = 6). (C) Fluorescence microscopy on pancreatic sections from Wt β−Tom and Tg7 β−Tom mice Red: tdTomato autofluorescence associated with β-cells. Scale bar: 40 μm. (D) Electron microscopy of 12w Wt and Tg7 islet preparations. Arrowhead indicates insulin granules. Arrows indicate the intercellular space between surrounding β-cells in Tg7 compared to control islets. Scale bar: 1 μm. Also shown is a zoom of the intercellular space between islet β-cells. (E) mRNA levels for ECM remodellers and collagen genes in islets from 12w Tg7 and Wt mice obtained by qPCR (n = 6). (F) Sirius red staining of pancreatic sections from 12w Tg7 mice and controls. (G) Collagen I content in islets from 12w Tg7 and Wt mice measured by ELISA. Islets from Tg7 mice were pre-treated with TGFβRI inhibitor Alk5i (1 uM) (n = 3–6). In panel G, one-way ANOVA with Sidak's multiple comparison test; otherwise, unpaired Student's t-test. Data are means ± SEM. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: The supernatant was collected, kept on ice, and processed using a Mouse Collagen Type I (COL1) ELISA kit (Abbexa, Cambridge, UK) according to the manufacturer's instructions.

Techniques: Western Blot, Fluorescence, Microscopy, Electron Microscopy, Control, Staining, Enzyme-linked Immunosorbent Assay, Comparison