Fig 2: Circ_0057558 inhibited AMPK signaling via targeting miR-206.A miR-206 levels in cells with the transfection of sh-circ_0057558 or sh-NC following FFA or control treatment. B ROCK1 mRNA levels in cells with the transfection of sh-circ_0057558 or sh-NC following FFA or control treatment. C Binding sites between the miR-206 and circ_0057558. D Relative luciferase activity of circ_0057558-WT and circ_0057558-MUT in cells with the transfection of miR-206 mimics or NC. E circ_0057558 and miR-206 levels following immunoprecipitation with anti-AGO2 or anti-IgG. F, G ROCK1/AMPK activities in cells with the transfection of sh-circ_0057558 or sh-NC following FFA or control treatment. H Protein levels of ROCK1 and p-AMPK in cells with the transfection of sh-circ_0057558 or sh-NC following FFA or control treatment. I Quantifications of H. *p < 0.05, **p < 0.01, and ***p < 0.001.

Fig 3: miR-206 inhibited lipogenesis and TG secretion in vivo.A Representative images of H&E staining (upper), oil-red o staining (middle), and IHC staining (bottom) in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. B Secreted TG levels in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. C miR-206 levels in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. D ROCK1 mRNA levels in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. E, F ROCK1/AMPK activities in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. G Protein levels of ROCK1, p-AMPK, and lipogenesis-related proteins in liver tissues from HFD-fed mice or control mice with injection of miR-206 mimics or NC. H Quantifications of G. *p < 0.05, **p < 0.01, and ***p < 0.001.

Fig 4: Effects of pro-oxidants on AMPK activity and Cn pathay. AMPKa (Thr172) activity in Raw264.7 and C2C12 cells were measured following treatment with DTPP + control or TPP + conjugated agents (MitoPQ and MitoMet) for 16 h at the indicated doses. A, Phosphorylated AMPKa level was measured in Raw264.7 cells and C2C12 cells. B, immunoblot analysis of phosphor-AMPKa protein along with GAPDH as loading control. The Bar diagram representing (the band intensity AMPKa Phos/tolal) The blots are representative of three different runs (n = 3). C-E, mRNA expression profile of different MtRS signaling markers at different time of hypoxia in HCT116 cells. The mRNA levels were monitored in qRT-PCR assays. The results from 3 independent experiments and normalized to beta actin mRNA levels and expressed as a fold change over the normoxic control. F, Matrigel invasion of control HCT116 cells treated with or without mito-targeted agents for 16h, before cells were loaded in to the chamber. G, quantitation of invading cells from triplicate experiments as in F. Significance was calculated by one-way ANOVA with Tukey's multiple-comparison test, and data are presented as treatment group versus no-treatment control. In all experiments, error bars represent standard deviations (*p < 0.05, **p < 0.01).

Fig 5: Effect of lut-7-O-rutinoside on mRNA expression levels of Lpl, Ucp-1, and PPARGC-1A (a) and protein levels of cAMP, chREBP, and AMPK (b) in L6 myotubes. Each value is presented as the mean ± SD for 6 replicates. *: p = 0.05 and **: p = 0.001, lut-7-O-rutinoside-treated L6 myotubes compared with control.