Fig. 3: Trametinib increases autophagic lysosomal activity in 5XFAD mice. | Molecular Psychiatry

Fig. 3: Trametinib increases autophagic lysosomal activity in 5XFAD mice.

From: MEK1/2 inhibition rescues neurodegeneration by TFEB-mediated activation of autophagic lysosomal function in a model of Alzheimer’s Disease

Fig. 3

A Analysis of the brain lysates from normal mice administered with trametinib for 2 weeks. Heatmap of RNA-Seq for 29 selected genes from autophagic lysosomal genes. The heatmap of RNA-Seq shows log10 FPKM (fragments per kilobase per million reads mapped) values for 29 selected genes (rows) and 6 samples (vehicle or trametinib administered, n = 3 in each group). Color corresponds to per-gene z-score that is calculated from log10 FPKM and represented using Morpheus heatmap program. B Representative western blot analysis of 5XFAD mice brain cortex lysates for indicated proteins. C Bars correspond to densitometric analysis of LC3II/α-tubulin. Data were presented as the mean ± S.E.M. One-way ANOVA followed by Dunnett’s post hoc analysis (F(2, 6) = 33.8, p = 0.0005; n = 3). D Bars correspond to densitometric analysis of p62. Data were presented as the mean ± S.E.M. One-way ANOVA followed by Dunnett’s post hoc analysis (F(2, 6) = 5.603, p = 0.0424; n = 3). E Bars correspond to densitometric analysis of cathepsin B. Data were presented as the mean ± S.E.M. One-way ANOVA followed by Dunnett’s post hoc analysis (F(2, 6) =  8.872, p = 0.0161; n = 3). F Immunofluorescence images of LC3, and LAMP1 in cortex layer V. Arrows indicate co-stained regions (n = 3 sagittal sections from each mouse, n = 3 mice per group). Scale bars, 10 μm. G Quantification of the co-stained ratio with LC3 and LAMP1. Data were presented as the mean ± S.E.M. Student’s t-test (p = 0.0381). H-K The 9-month-old 5XFAD mice were administered either vehicle or 0.1 mg/kg of trametinib for 6 weeks by oral gavage once a day. Immunofluorescence staining images of Aβ, LAMP1, and MAP2 in the cortex of 5XFAD mice (H). Scale bars, 50 μm. Quantification of LAMP1 intensity within cell body of MAP2 + neurons (I) (F(2, 38) = 4.320, p = 0.0204). Immunofluorescence staining images of Aβ, LAMP1, and SMI31 in the cortex of 5XFAD mice (J). Scale bars, 50  μm. Quantification of the area of dystrophic axon within plaque (K) (F(2, 24) = 46.67, p < 0.0001). Data were presented as the mean ± S.E.M. One-way ANOVA followed by Dunnett’s post hoc analysis. *p < 0.05; **p < 0.01; ***p < 0.001 versus 5XFAD-Veh group.

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