Figure 3. High expression of GSK-3β and Tau404 in injured HT-22 cells and diabetic hippocampal tissues. (A) Immunostaining analysis showing the enhanced colocalized relationship in Con, DM, and DE groups, progressively (as evidenced by overplay yellow areas in Merge image). Also, GSK-3β and Tau 404 Con are highly expressed in DM, and DE groups, progressively. (B, C) The immunoblot suggesting a high expression of GSK-3β, GSK-3β(T216), and Tau 404 in Con, DM, and DE groups, progressively. However, the expression levels of Tau 396 and Tau 404 are not different between DM and DE groups. (D, F) In vitro experiment (mimicking) which demonstrates higher expression levels of GSK-3β in Glu, AGEs, and H2O2 groups, vs Con in HT-22 cells. (E, G) The IP analysis demonstrating a binding relationship between GSK-3β and Tau 404, and this relationship is enhanced in DM and DE groups. *P < 0.05 vs. control group, #P < 0.05 vs. the DM group, n =3 for each group. For Figure 3F, *, **, and *** means P < 0.05, < 0.01, < 0.001 vs Con; bar = 100μm.