Fig. 3

TWD induces impaired Akt-Gsk3β signaling in the brain. a Western blot analysis of hippocampal lysates showing decreased phosphorylation of S473 in Akt1 in mice with TWD as compared to STD (p < 0.001). Diet had no effect on total Akt1 levels. b Similarly, phosphorylation of S474 in Akt2 is decreased in mice with TWD as compared to STD (p < 0.01). Diet had a minor, but statistically significant effect on total Akt2 levels (p < 0.05). c Also, phosphorylation of T308/309/305 residue in Akt1/2/3, respectively, was significantly decreased in mice with TWD as compared to STD (p < 0.001). Total Akt1/2/3 levels showed no significant changes. d Phosphorylation of Gsk3β at the inhibitory S9 residue was significantly decreased in mice with TWD as compared to mice with STD (p < 0.01). A+ transgene (APPswe/PS1dE9) increased slightly, but significantly Gsk3β S9 phosphorylation (p < 0.05) as well as total Gsk3β levels (p < 0.05). e A representative Western blot image and quantification of hippocampal lysates showing increased levels of the autophagy markers p62 and LC3B in TWD mice as compared to STD mice (p < 0.05). Phosphorylated protein levels were normalized to their respective total protein levels in cell lysates and total protein levels were normalized to Gapdh or β-actin. All results are shown as mean + SEM, n = 5–7 mice/group, Two-way ANOVA. AwTw = wild-type, AwT+ = Tau P301L, A+Tw = APPswe/PS1dE9, A+T+ = APPswe/PS1dE9 x Tau P301L