Fig 1: The downregulation of miR-133a-3p and miR-1a-3p promotes skeletal muscle growth by activating Akt/mTOR signaling. (A) Potential binding sites for miR-133a-3p and miR-1a-3p in the 3'UTR of Insr and EIF4E mRNA, respectively, predicted by RNAhybrid (MFE: minimal free energy); (B) the relative expression of the miRNA target gene after GAA treatment or miRNA mimic overexpression (Student’s t-test) (R: Pearson correlation coefficient); (C) the relative expression of the miRNA target gene after miRNA mimic overexpression (Tukey’s post-hoc test); (D) luciferase reporter assay indicated that transfection with miRNA mimic significantly suppressed the relative activity of luciferase (Student’s t-test); (E,F) the Akt, mTOR and S6K phosphorylation levels detected by western blotting after the overexpression of miRNA mimic or GAA treatment (Tukey’s post-hoc test). The results are expressed as mean ± S.D. of three independent experiments. * p < 0.05; ** p < 0.01 compared with negative control (on the bars) or mimic control (on the bars) or between the indicated groups.
Fig 2: mTOR signaling pathway analysis. (A) The green and yellow areas indicate hippocampus and hypothalamus respectively. (B) The red area indicates amygdala. (C) The blue area indicates prefrontal cortex. (D) Representative Western blots of hippocampus, hypothalamus, amygdala and prefrontal cortex. The expression of AKT (E and F), p-AKT (G and H), p70S6K (K and L), p-mTOR (O and P) and mTOR (M and N) in hippocampus, hypothalamus, amygdala and prefrontal cortex. The ratios of p-AKT/AKT and p-mTOR/mTOR in hippocampus, hypothalamus amygdala and prefrontal cortex (I, J, Q, R). *P < 0.05; **P < 0.01; ***P<0.001; ****P < 0.0001 vs sham. n=3 mice per group. Data are presented as mean±SEM. One-way ANOVA was used for comparison among groups.
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