Figure 5: miR-24 Regulates OGT and ATG4A to Play Cardioprotective Effects

Sequence alignment of miR-24 targeting sites on OGT (A) and ATG4A (B) 3ʹUTR among mammals based on the Targetscan database information. (C) miR-24 inhibited luciferase activity of WT 3ʹUTR of OGT, ATG4A, and BIM, but exhibited no effect on luciferase activity of each mutant 3ʹUTR. The assays were performed in triplicate from 3 independent experiments. (D) The change in intracellular miR-24 levels when cardiomyocytes were transfected with miR-24 mimics and performed in triplicate from 3 independent experiments. (E) (Top) Representative Western blot analyses of OGT, ATG4A, and BIM expression in response to miR-24 overexpression in cardiomyocytes, normalized to HSP90 and actin. (Bottom) Cardiomyocytes were transfected with miR-24 mimic (Ambion mirVana miRNA mimic) or negative control via Lipofectamine RNAiMAX Transfection Reagent. A total of 48 h after transfection, cells were treated with 100 nmol/l insulin for another 1 h before harvested. The expression of OGT was analyzed by western blot. (F) OGT expression and O-GlcNAcylation of heart lysates from WT mice or mice with cardiomyocyte-specific overexpression of miR-24, and heart lysates from in db/db mice with control or miR-24 mimic (Life Technologies, 5 mg/kg) delivery. Abbreviations as in Figure 1.



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