Research Paper Volume 15, Issue 4 pp 932—946

Figure 5. Investigation of signaling pathways associated with AMPKα isoform inhibition in aged mice revealed de-phosphorylation of eIF2α in the hippocampus of the AMPKα1 cKO mice. (A) Western blot experiments showed no alterations in phosphorylation levels of eEF2 (Thr56), mTOR (Ser2448), and AKT (Ser473) in hippocampal synaptosome lysate of AMPKα1 cKO or AMPKα2 cKO mice, compared to the Cre+/- mice. Representative Western blot gels and quantification data presented in bar graphs are shown. n= 4-6 per group. p>0.05, One-way ANOVA. (B) Levels of phospho-eIF2α (Ser51) were decreased in hippocampal synaptosome lysate of AMPKα1 cKO mice, compared to Cre+/- or AMPKα2 cKO mice group. n=7-8 per group. *p<0.05, **p<0.01, One-way ANOVA and Tukey’s test. (C, D) Levels of ATF4 and PERK were not altered in either AMPKα1 cKO or AMPKα2 cKO mice. n=5-6 per group. p>0.05, One-way ANOVA. (E) Representative images and quantification from the SUnSET de novo protein synthesis assay. n=3-4. p>0.05, One-way ANOVA. (F) Hippocampal polyribosome formation was unaltered in AMPKα1 cKO or AMPKα2 cKO mice, compared to the Cre+/- group. Representative transmission electron microscopy (TEM) images of hippocampal CA1 and cumulative data of polyribosome quantification were shown. Polyribosomes were indicated with red arrows. n=3 mice per group (8-10 ROI measurements per mouse). p=0.43. One-way ANOVA.