miR-27b-3p Attenuates Muscle Atrophy by Targeting Cbl-b in Skeletal Muscles

Figures

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  Figure 1. miRNA-seq analysis for the Dex group and Con group. (A) miRNA array shows miRNA dysregulation from Dex-induced models. (B) Volcano plot of DEMs from Dex VS Con.

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  Figure 2. mRNA-seq analysis for the Dex group and Con group. (A) Volcano plot of DEGs from Dex VS Con. (B) GO enrichment analysis of DEGs. (C) Classification analysis of the KEGG pathway of DEGs.

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  Figure 3. DEMs–DEGs interactive network. (A) Interaction network between upregulated DEMs and downregulated DEGs. (B) The interaction network between downregulated DEMs and upregulated DEGs.

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  Figure 4. miRNA-27b-3p can affect muscle atrophy marker genes and target Cbl-b. (A) miR-27b-3p can relieve Dex-induced atrophy in myotubes. (B,C) Sequence of miR-27b-3p binding site with target gene. (D) The luciferase assay was performed by co-transfection of wild-type or mutant Cbl-b 3′UTR with miR-27b-3p mimic or mimic NC in C2C12 cells. (E,F) After transfection with miR-27b-3p mimic or inhibitor, Cbl-b protein expression was obtained. Results are shown as mean ± S.E.M. and the data are representative of at least three independent assays. Independent sample t-test was used to analyze the statistical differences between groups. (* p < 0.05; ** p < 0.01).

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  Figure 5. Some differences between the Dex group and Con group mice. (A) Differences in body weight, gastrocnemius weight, tibial anterior muscle weight, and morphology between the two groups of Kunming mice. (B) Diameter distribution of the gastrocnemius tube in the two groups. (C) Diameter distribution of the tibial anterior muscle tube in the two groups. (D) Differential expression of Cbl-b, Atrogin-1, Murf-1, FoxO1, and FoxO3 in the two groups of mice. Dex, dexamethasone. Con, control. Results are shown as mean ± S.E.M. and the data are representative of at least three independent assays. Independent sample t-test was used to analyze the statistical differences between groups. (* p < 0.05; ** p < 0.01; *** p < 0.001).

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  Figure 6. Cbl-b affects muscle atrophy. (A) Relative gene expression after the overexpression of Cbl-b. (B) Relative gene expression after losing function of Cbl-b. (C) Relative protein level after the overexpression of Cbl-b. (D) Relative gene expression after losing function of Cbl-b in induced muscle atrophy cells. Results are shown as mean ± S.E.M. and the data are representative of at least three independent assays. Independent sample t-test was used to analyze the statistical differences be-tween groups. (* p < 0.05; ** p < 0.01; *** p < 0.001).