Fig. 2
Ex-4 reduces tau hyperphosphorylation by increasing brain-derived insulin via activating Wnt/β-catenin pathway in HF-diabetic mice. A Workflow of the animal study for C57BL/6 mice. Insulin levels B and c-peptide levels C in CSF were measured by ELISA kits (n = 6/group). D Represented immunofluorescence staining coronal brain sections of each group mice were performed for insulin (green) and DAPI (blue). Scale bar = 100 μm. E mRNA levels of Ins2 in the hippocampus were measured by RT-qPCR assay. β-actin as the internal control. (n = 5/group). F The levels of the phosphorylated tau at Ser199, Ser202, Ser396, and Thr217 sites and total tau in each group; the activation of Wnt/β-catenin pathway as evidenced by the levels of the np-β-catenin to total β-catenin; insulin signalling activation as P-AKTS473 to total AKT and P-GSK-3βS9 to total GSK-3β were examined through Western blot analysis, (n = 5/group). G The gray density of E. H Represented immunohistochemical staining of coronal brain sections from mice of each group was performed with the antibody against phosphorylated tau at Thr231. Scale bar = 100 μm. For D and H, HIP: hippocampus, CA: cornue ammonis, DG: dentate gyrus. Results are representative of three independent experiments. Values are presented as mean ± SD. **P < 0.01, ***P < 0.001, ****P < 0.0001. vs. CTL; ††P < 0.01, †††P < 0.001, ††††P < 0.0001. vs. HF-diabetic; ‡‡P < 0.01, ‡‡‡P < 0.001, ‡‡‡‡P < 0.0001. vs. Ex-4