Fig. 6.

BMAL1 overexpression-induced cognitive improvement depends on PI3K/Akt/GSK-3β signaling pathway in db/db mice. A The experimental timeline. B, C The total distance traveled and average velocity was assessed in the open field test (n = 8 per group). D, E The duration and frequency of exploring the novel object were assessed in the novel object recognition test (n = 8 per group). F, G Acquisition errors and probe errors of the Barnes maze test were assessed for spatial learning ability (n = 8 per group). H Representative track images of mice in the probe trial in the Barnes maze test. I Western blots of the phosphorylation levels of PI3K, Akt, and GSK-3β in the hippocampus (n = 3 per group). J Western blots of the phosphorylation levels of tau at Ser396, Thr231, and Ser199 sites in the hippocampus (n = 3 per group). K Western blots of the expression levels of APP and BACE1 in the hippocampus (n = 3 per group). L Immunohistochemical for Aβ plaques in the hippocampus in four groups (Scale bars = 400 µm in low-magnification and scale bars = 100 µm in high-magnification image) (n = 3 per group). M ELISA of Aβ1-40 and Aβ1-42 in the hippocampus (n = 4 per group). Data were plotted as means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. The data were analyzed by two-way ANOVA followed by post hoc Tukey’s HSD test