Journal: Military Medical Research

Article Title: USP18 exacerbates myocardial I/R injury by inhibiting Parkin mitophagy through the deubiquitinase PTEN-L

doi: 10.1016/j.mmr.2026.100004

Figure Lengend Snippet: USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated protein 1 light chain 3; VDAC. Voltage-dependent anion channel.

Article Snippet: To block mitophagy, the selective dynamin-related protein 1 (Drp1) inhibitor Mdivi-1 was used (50 μmol/L, MedChemExpress, USA).

Techniques: Inhibition, Electron Microscopy, Transfection, Infection, Ubiquitin Proteomics, Knock-Out

Journal: The American Journal of Pathology

Article Title: The D2.B10- Dmd mdx /J Mouse Model of Duchenne Muscular Dystrophy Exhibits a Severe Mitochondrial Deficiency Not Observed in the C57BL/10ScSn- Dmd mdx /J Mouse

doi: 10.1016/j.ajpath.2025.09.005

Figure Lengend Snippet: Relative protein expression changes to the latent transforming growth factor (TGF)-β–binding protein 4 (LTBP4) pathway and annexin A6 (ANXA6). LTBP4 is a known genetic modifier in human patients with Duchenne muscular dystrophy and impacts TGF-β signaling. TGF-β acts on SMAD4, which directly interacts with mitochondrial cytochrome c oxidase subunit 2 (MTCO2) and can cause mitochondrial fragmentation via dynamin-related protein 1 (DRP1). A – L: All proteins were assessed for expression levels ( A – F ) relative to total protein ( G – L ). H: TGF-β levels were elevated in D2-WT mice compared with B10-WT mice, and complex IV subunit 2 (MTCO2) was elevated in the D2- mdx mice compared with D2-WT. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Antibodies against latent TGF-β–binding protein 4 (LTBP4; Novus Biologicals, Centennial, CO; NBP2-43671; 1:500), TGF-β (Novus Biologicals; NBP2-46108; 1:2000), annexin A6 (ANXA6; ProteinTech, Rosemont, IL; 68086-1-Ig; 1:2000), mitochondrial cytochrome c oxidase subunit 2 (MTCO2; Thermo Fisher; A-6404; 1:500), SMAD4 (ProteinTech; 10231-1-AP; 1:100), dynamin-related protein 1 (DRP1; ProteinTech; 12957-1-AP; 1:1000), and Total OXPHOS Rodent WB Antibody Cocktail (Abcam; ab110413; 1:25,000) were used.

Techniques: Expressing, Binding Assay