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: eLife

Article Title: The dual molecular identity of vestibular kinocilia bridges structural and functional traits of primary and motile cilia

doi: 10.7554/eLife.108071

Figure Lengend Snippet: ( A ) Violin plots of the expression of 72 genes in four different HC types. ( B ) Validation of differential expressions of nine genes (with underline in A ) in cochlear and vestibular HCs in thin section. Bar: 10 μm for all images in B. ( C ) Confocal images of expression of DNM1, SLC7A14, and TJAP1 in cochlear and vestibular HCs. Bar: 10 μm for all images in C.

Article Snippet: Primary antibodies against DNM1 (NBP2-48950, Novus Biologicals), SLC7A14 (HPA045929, Sigma), TJAP1 (NBP1-80902, Novus), FOXJ1 (14-9965-82, Thermo Fisher), CCDC39 (HPA035364, Sigma), CCDC40 (PA5-54653, Thermo Fisher), DNAH5 (31079-1-AP, Thermo Fisher), DNAH6 (HPA036391, Sigma), TEKT1 (HPA044444, Millipore Sigma), CLUAP1 (PA5-83710, Thermo Fisher), IFT172 (28441-1-AP), and acetylated tubulin (T6793, Sigma) were incubated with the tissues for 12 hr at 4°C.

Techniques: Expressing, Biomarker Discovery

Journal: bioRxiv

Article Title: GM-CSF and M-CSF Driven Differentiation Differentially Regulates Chikungunya Virus Infection and Antiviral Responses in Human Monocyte-Derived Macrophages

doi: 10.64898/2026.03.11.710213

Figure Lengend Snippet: (A) Schematic representation of compounds targeting CHIKV entry and internalization mechanisms in macrophages. GM-Mϕ cultured on 96-wells plates were incubated with macrophage media containing compounds inhibiting phagocytosis: (B) cytochalasin D, macropinocytosis: (C) EIPA, clathrin: (D) pitstop2, dynamin: (E) dynasore, or endocytosis/viral fusion: (F) bafilomycin A1, (H) chloroquine, (G) ammonium chloride, or vehicle control: DMSO for 2 hours at 37°C. Cells were then infected with CHIKV 181/25 (MOI=1.0) for 1 hour 37°C, and re-supplemented with media containing compound for a total incubation time of 24 hours. CHIKV infection was evaluated via immunofluorescence by staining for expression of CHIKV capsid protein and total cell count was determined by DAPI staining. Percent infection was determined by determining number of infected cells over total cells. Cell viability was determined via MTT assay. Data for percent (%) infection and cell viability of compounds is represented as normalized to DMSO vehicle control. (I) IC50 and CC50 values and corresponding dose-response curve for viral infectivity and cell viability was determined via non-linear [inhibitor] vs normalized response – variable non-linear slope analysis. Data represented as means ± SEM. Antiviral and cell viability assays were performed in technical triplicate (n=3 donors).

Article Snippet: Other compounds targeting micropinocytosis (EIPA), clathrin (pitstop 2), and dynamin (dynasore) were obtained from MedChemExpress.

Techniques: Cell Culture, Incubation, Control, Infection, Immunofluorescence, Staining, Expressing, Cell Characterization, MTT Assay