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: Journal of Advanced Research

Article Title: OGT regulates histone demethylation and alleviates osteoarthritis progression by O-GlcNAcylation of KDM6B

doi: 10.1016/j.jare.2026.01.003

Figure Lengend Snippet: OGT is downregulated in cartilage from OA patients. (A) Differentially expressed genes in cartilage from control individuals and OA patients. (B) Volcano plot showing upregulated and downregulated genes in OA cartilage. (C) Relative expression levels of OGT in cartilage from control and OA patients. (D) GSEA demonstrating protein O-linked glycosylation enriched in OA cartilage (NES = -1.53). (E) Representative X-ray and cartilage histology images from OA patients. (F, G) Western blot analysis and quantification of OGT in intact and damaged cartilage (n = 6). (H) Representative H&E and Safranin O/Fast Green (SO&FG) staining of intact and damaged cartilage (n = 3. Scale bars, 1000 μm). (I) Representative immunofluorescence images of OGT in intact and damaged cartilage (n = 3. Scale bars, 100 μm). (J) Representative H&E and SO&FG staining of human cartilage treated with IL-1β (10 ng/mL) (n = 3. Scale bars, 500 μm). (K) Representative immunofluorescence images of OGT in explants treated with IL-1β (10 ng/mL) (n = 3. Scale bars, 100 μm). (L, M) Quantification of positive cells in panels G and H (n = 3). Data are presented as the means ± SEM. P values are from two-tailed unpaired Student’s t -test. *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: The cartilage was cut into small pieces, washed three times with sterile phosphate-buffered saline (PBS), and cultured in DMEM/F-12 medium supplemented with 10 % FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin for 24 h. Subsequently, the explants were cultured for an additional 7 days in medium supplemented with 10 ng/mL recombinant human IL-1β (Abclonal, RP00002).

Techniques: Control, Expressing, Glycoproteomics, Western Blot, Staining, Immunofluorescence, Two Tailed Test