Review

cytochrome c oxidase subunit 4 (Cell Signaling Technology Inc)

Cell Signaling Technology Inc is a verified supplier

Cell Signaling Technology Inc manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Cell Signaling Technology Inc cytochrome c oxidase subunit 4
Cytochrome C Oxidase Subunit 4, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse/COX+IV+(4D11-B3-E8)+Mouse+mAb/pmc12816905-99-54-60
Average 96 stars, based on 221 article reviews

cytochrome c oxidase subunit 4 - by Bioz Stars, 2026-09

96/100 stars

Images

other:

Article Title: Testosterone Exposure During Fetal Masculinization Programming Window Determines the Kidney Size in Adult Mice
Article Snippet: anti‐Phosphohistone H3 , Mouse , Cell Signaling Technology , 9706 , AB_331748 , 1:200 , IF.

Article Title: Picalm coordinates clathrin-mediated endocytosis and actin remodeling during myogenesis
Article Snippet: Akt , Monoclonal , Mouse , Cell signaling , 2920.

Immunohistochemistry:

Article Title: AAV.PHP.eB-based strategies for precise modulation of α7 nicotinic acetylcholine receptor in neurons and astrocytes in the adult mouse brain.
Article Snippet: .. Immunohistochemistry was performed on free-floating sections by using overnight at 4°C the following antibodies: mouse anti GFAP antibody (#3670, 1:300, Cell Signaling) and anti-mouse NeuN (MAB377, 1:300, Millipore). .. The fluorescent secondary antibody Alexa Fluor 488 donkey anti-mouse (A32723, Invitrogen; 1:500) was then used for 90 minutes at RT.

Paraffin Section:

Article Title: Lymphocyte Antigen 6G Mediates Vagotomy‐Associated Reduction in Body Weight
Article Snippet: Antigen retrieval was performed using the antigen retrieval solution (DIVA Decloaker, pH 6, #DV2004, BiocareMedical; 10×, diluted in MilliQ). .. Sections were blocked in 5% horse serum in 0.05% TBST for 30 min at RT. eWAT paraffin section were stained with rabbit anti‐mouse anti‐perilipin 1 (Cell signaling technology, #9349 T) and rat anti‐mouse anti‐Ly6G antibody (BD Biosciences, #551459), washed with 0.05% TBST, and incubated with secondary antibodies (horse anti‐rabbit IgG Antibody (H + L), DyLight 488 (Vector Laboratories, #DI‐1088), and Goat Anti‐Rat IgG Antibody (H + L) DyLight 594 (Abcam, #ab98422)) or horse anti‐rabbit IgG antibody (H+L), DyLight 594 (Vector Laboratories, #DI‐1094). .. Nuclei were stained with DAPI (4′,6‐diamidino‐2‐phenylindole), and samples were mounted with Dako mounting media (Agilent).

Staining:

Incubation:

Article Title: Distinct cMET inhibitors uncover pharmacological heterogeneity in SHH medulloblastoma cell lines.
Article Snippet: Interphase duration, mitotic time and time of cell death were recorded and plotted using GraphPad Prism 8. .. Blots were blocked in 5% milk, PBS-Tween 0.1% solution and incubated with primary antibodies overnight at 4 C followed by one hour incubation with HRP-conjugated secondary antibodies raised in rabbit or mouse (anti-Rabbit IgG from Cell Signalling Technologies, 1:5000; anti-Mouse IgG from GE Healthcare, 1:3000). .. Band visualisation was completed by using West Femto Chemilumiscent Substrate (ThermoFisher, UK) and images were recorded by using a G: BOX SynGene system.

Article Title: Double-wall carbon nanotubes promote ASC speck formation and inflammasome activation in vascular endothelial cells.
Article Snippet: Carbon nanotubes (CNTs) are important industrial nanomaterials, yet their manufacture presents potential risks for respiratory exposure among workers.. CNTs have a high aspect ratio, which raises concerns that they may cause toxicity similar to that of asbestos.. Asbestos is known to activate the NLR family pyrin domain containing 3: NLRP3, a key component of inflammasome.

Western Blot:

Article Title: Testosterone Exposure During Fetal Masculinization Programming Window Determines the Kidney Size in Adult Mice
Article Snippet: anti‐Phospho‐AKT (ser473) , Rabbit , Cell Signaling Technology , 9271 , AB_329825 , 1:1000 , WB. .. anti‐Phospho‐S6 ribosomal protein (Ser235/236) , Mouse , Cell Signaling Technology , 2211 , AB_331679 , 1:1000 , WB. .. Anti‐Beta‐Actin , Mouse , Sigma‐Aldrich , A1978 , AB_476692 , 1:2500 , WB.

Image Search Results

USP18 expression is induced by cardiac I/R injury. a Gene Ontology (GO) enrichment analysis of the RNA-seq data in I/R and control hearts at 24 h postsurgery ( n =3). b Heatmap displaying differentially expressed ubiquitination-associated genes in I/R and control hearts at 24 h postsurgery ( n =3). c Protein levels of USP18 in donor heart tissue and patients with ischemic heart disease (IHD) ( n =4). d Immunofluorescence staining of USP18 and α-actin in donor heart tissue and patients with IHD ( n =3). Scale bar=35 μm (left) and 50 μm (right). e Protein levels of USP18 in heart tissue post-IR ( n =4). f Immunofluorescence staining of USP18 and WGA in heart tissue 24 h after I/R injury ( n =5). Scale bar=50 μm. g Protein expression of USP18 in primary neonatal rat cardiomyocytes (NRVMs) after H/R challenge ( n =4). h Immunofluorescence staining of USP18 in cardiomyocytes after H/R challenge. Scale bar=50 μm. ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WGA. Wheat germ agglutinin; DAPI. 4′,6-diamidino-2-phenylindole; H/R. Hypoxia/reoxygenation; RNA-seq. RNA sequencing; MAPK. Mitogen-activated protein kinase; GABA. γ-aminobutyric acid.

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 expression is induced by cardiac I/R injury. a Gene Ontology (GO) enrichment analysis of the RNA-seq data in I/R and control hearts at 24 h postsurgery ( n =3). b Heatmap displaying differentially expressed ubiquitination-associated genes in I/R and control hearts at 24 h postsurgery ( n =3). c Protein levels of USP18 in donor heart tissue and patients with ischemic heart disease (IHD) ( n =4). d Immunofluorescence staining of USP18 and α-actin in donor heart tissue and patients with IHD ( n =3). Scale bar=35 μm (left) and 50 μm (right). e Protein levels of USP18 in heart tissue post-IR ( n =4). f Immunofluorescence staining of USP18 and WGA in heart tissue 24 h after I/R injury ( n =5). Scale bar=50 μm. g Protein expression of USP18 in primary neonatal rat cardiomyocytes (NRVMs) after H/R challenge ( n =4). h Immunofluorescence staining of USP18 in cardiomyocytes after H/R challenge. Scale bar=50 μm. ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WGA. Wheat germ agglutinin; DAPI. 4′,6-diamidino-2-phenylindole; H/R. Hypoxia/reoxygenation; RNA-seq. RNA sequencing; MAPK. Mitogen-activated protein kinase; GABA. γ-aminobutyric acid.

Article Snippet: To establish a cardiac-specific USP18 knockout mice model (USP18-cKO), USP18 flox/flox mice (C57BL/6 J; Shanghai Model Organisms, NM-cKO-215042) were bred with α-MHC-MerCreMer mice (C57BL/6 J; Jackson Laboratory, stock No. 024992).

Techniques: Expressing, RNA Sequencing, Control, Ubiquitin Proteomics, Immunofluorescence, Staining

$USP18 deficiency mitigates mitochondrial dysfunction, acute cardiac I/R injury, and cardiac remodeling in vivo. a 2,3,5-triphenyltetrazolium chloride (TTC) staining of heart tissue 24 h post-I/R in WT and I/R groups ( n= 5). Scale bar=0.5 cm. ⁎⁎⁎ P <0.001. b Mitochondrial DNA ( n =6) and mitochondrial complexes I and II–III activity ( n =5) 24 h post-I/R in each group. ⁎⁎ P <0.01, ⁎⁎⁎⁎ P <0.0001. c Representative electron micros c opy images of heart sections 24 h post-I/R; quantitative analysis of the mitochondrial volume density and percent of mitochondria with cristae loss in each group ( n= 5). Scale bar=10 μm (top) and 6 μm (bottom). Red arrows indicate mitochondria with cristae loss ⁎⁎⁎ P <0.001. d Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in mitochondria in the indicated groups ( n= 4). ⁎⁎ P <0.01, ⁎⁎⁎⁎ P <0.0001, ns non-significant. e H&E staining, PSR staining, and quantitative statistical analysis of cell size and left ventricle (LV) fibrotic area in WT mice and USP18-cKO mice at 4 weeks after I/R injury ( n= 5). Scale bar=1 mm (left), 50 μm (middle), and 100 μm (right). ⁎⁎ P <0.01. f Representative B-mode and M-mode echocardiographic images of LV from WT mice or USP18-cKO mice 4 weeks after I/R injury. g Cardiac function of WT mice and USP18-cKO mice after I/R injury at the indicated time points ( n= 6). ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001 vs . WT, ns non-significant. USP18 . Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; ATP. Adenosine triphosphate; H&E. Hematoxylin and eosin; PSR. Picrosirius red; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; USP18-cKO. Ubiquitin-specific protease 18 conditional knockout; LVIDd. Left ventricle internal diameter at diastole; LVISd. Left ventricle internal diameter at systole; LVEF. Left ventricle ejection fraction; LVFS. Left ventricle fractional shortening.$

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 deficiency mitigates mitochondrial dysfunction, acute cardiac I/R injury, and cardiac remodeling in vivo. a 2,3,5-triphenyltetrazolium chloride (TTC) staining of heart tissue 24 h post-I/R in WT and I/R groups ( n= 5). Scale bar=0.5 cm. ⁎⁎⁎ P <0.001. b Mitochondrial DNA ( n =6) and mitochondrial complexes I and II–III activity ( n =5) 24 h post-I/R in each group. ⁎⁎ P <0.01, ⁎⁎⁎⁎ P <0.0001. c Representative electron micros c opy images of heart sections 24 h post-I/R; quantitative analysis of the mitochondrial volume density and percent of mitochondria with cristae loss in each group ( n= 5). Scale bar=10 μm (top) and 6 μm (bottom). Red arrows indicate mitochondria with cristae loss ⁎⁎⁎ P <0.001. d Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in mitochondria in the indicated groups ( n= 4). ⁎⁎ P <0.01, ⁎⁎⁎⁎ P <0.0001, ns non-significant. e H&E staining, PSR staining, and quantitative statistical analysis of cell size and left ventricle (LV) fibrotic area in WT mice and USP18-cKO mice at 4 weeks after I/R injury ( n= 5). Scale bar=1 mm (left), 50 μm (middle), and 100 μm (right). ⁎⁎ P <0.01. f Representative B-mode and M-mode echocardiographic images of LV from WT mice or USP18-cKO mice 4 weeks after I/R injury. g Cardiac function of WT mice and USP18-cKO mice after I/R injury at the indicated time points ( n= 6). ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001 vs . WT, ns non-significant. USP18 . Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; ATP. Adenosine triphosphate; H&E. Hematoxylin and eosin; PSR. Picrosirius red; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; USP18-cKO. Ubiquitin-specific protease 18 conditional knockout; LVIDd. Left ventricle internal diameter at diastole; LVISd. Left ventricle internal diameter at systole; LVEF. Left ventricle ejection fraction; LVFS. Left ventricle fractional shortening.

Techniques: In Vivo, Staining, Activity Assay, Ubiquitin Proteomics, Knock-Out

$Overexpression of USP18 in the heart exacerbates mitochondrial dysfunction, acute cardiac injury, and cardiac remodeling following I/R in mice. a TTC staining of heart tissue 24 h post-I/R in each group ( n= 5). Scale bar=0.5 cm. ⁎⁎⁎ P <0.001. b Mitochondrial DNA levels ( n= 6) and mitochondrial complexes I and II–III activity ( n= 5) 24 h post-I/R in each group. ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. c Representative electron microscopy images of heart sections 24 h post-I/R. Quantitative analysis of mitochondrial volume density and the percent of mitochondria with cristae loss in each group ( n= 5). Scale bar=10 μm (top) and 6 μm (bottom). Red arrows indicate mitochondria with cristae loss. ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. d Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in mitochondria in the indicated groups ( n= 4). ⁎ P <0.05, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. e H&E staining, PSR staining, and quantitative statistical analysis of cell size and left ventricl e (LV) fibrotic area in AAV9-USP18-transfected mice at 4 weeks after I/R injury ( n= 5). Scale bar=1 mm (left), 50 μm (middle), and 100 μm (right). ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. f Representative B-mode and M-mode echocardiographic images of LV from AAV9-USP18-transfected mice 4 weeks after I/R injury. g Cardiac function of AAV9-USP18-transfected mice after I/R injury at the indicated time points ( n= 6). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001 vs . AAV9-NC, ns non-significant. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; NC. Negative control; AAV9. Adeno-associated virus serotype 9; AAV9-USP18. Adeno-associated virus serotype 9 encoding USP18; TTC. 2,3,5-triphenyltetrazolium chloride; ATP. Adenosine triphosphate; H&E. Hematoxylin and eosin; PSR. picrosirius red; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; LVIDd. Left ventricle internal diameter at diastole; LVISd. Left ventricle internal diameter at systole; LVEF. Left ventricle ejection fraction; LVFS. Left ventricle fractional shortening.$

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: Overexpression of USP18 in the heart exacerbates mitochondrial dysfunction, acute cardiac injury, and cardiac remodeling following I/R in mice. a TTC staining of heart tissue 24 h post-I/R in each group ( n= 5). Scale bar=0.5 cm. ⁎⁎⁎ P <0.001. b Mitochondrial DNA levels ( n= 6) and mitochondrial complexes I and II–III activity ( n= 5) 24 h post-I/R in each group. ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. c Representative electron microscopy images of heart sections 24 h post-I/R. Quantitative analysis of mitochondrial volume density and the percent of mitochondria with cristae loss in each group ( n= 5). Scale bar=10 μm (top) and 6 μm (bottom). Red arrows indicate mitochondria with cristae loss. ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. d Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in mitochondria in the indicated groups ( n= 4). ⁎ P <0.05, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. e H&E staining, PSR staining, and quantitative statistical analysis of cell size and left ventricl e (LV) fibrotic area in AAV9-USP18-transfected mice at 4 weeks after I/R injury ( n= 5). Scale bar=1 mm (left), 50 μm (middle), and 100 μm (right). ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. f Representative B-mode and M-mode echocardiographic images of LV from AAV9-USP18-transfected mice 4 weeks after I/R injury. g Cardiac function of AAV9-USP18-transfected mice after I/R injury at the indicated time points ( n= 6). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001 vs . AAV9-NC, ns non-significant. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; NC. Negative control; AAV9. Adeno-associated virus serotype 9; AAV9-USP18. Adeno-associated virus serotype 9 encoding USP18; TTC. 2,3,5-triphenyltetrazolium chloride; ATP. Adenosine triphosphate; H&E. Hematoxylin and eosin; PSR. picrosirius red; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; LVIDd. Left ventricle internal diameter at diastole; LVISd. Left ventricle internal diameter at systole; LVEF. Left ventricle ejection fraction; LVFS. Left ventricle fractional shortening.

Techniques: Over Expression, Staining, Activity Assay, Electron Microscopy, Transfection, Ubiquitin Proteomics, Negative Control, Virus

Effects of USP18 on H/R-induced injury to NRVMs. a-d NRVMs were transfected with USP18 siRNA and exposed to H/R. Scale bar=26 μm (top) and 50 μm (bottom). Representative confocal microscopy images showing mitochondrial morphology probed with MitoTracker Red ( n= 6) and quantification of mitochondrial length, mitochondrial fragmentation, and mitochondrial DNA levels ( n= 6) ( a and b ). Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in NRVMs in the indicated groups ( n= 4) ( c and d ). e-h NRVMs were infected with Ad-USP18 and exposed to H/R. Scale bar=26 μm (top) and 50 μm (bottom). Representative confocal microscopy images showing mitochondrial morphology probed with MitoTracker Red ( n= 6) and quantification of mitochondrial length, mitochondrial fragmentation, and mitochondrial DNA levels ( n= 6) ( e and f ). Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in NRVMs in the indicated groups ( n= 4) ( g and h ). ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001, ns non-significant. US P 18. Ubiquitin-specific protease 18; H/R. Hypoxia/reoxygenation; NC. Negative control; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; NRVMs. Neonatal rat ventricular cardiomyocytes; ATP. Adenosine triphosphate; OCR. Oxygen consumption rate.

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: Effects of USP18 on H/R-induced injury to NRVMs. a-d NRVMs were transfected with USP18 siRNA and exposed to H/R. Scale bar=26 μm (top) and 50 μm (bottom). Representative confocal microscopy images showing mitochondrial morphology probed with MitoTracker Red ( n= 6) and quantification of mitochondrial length, mitochondrial fragmentation, and mitochondrial DNA levels ( n= 6) ( a and b ). Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in NRVMs in the indicated groups ( n= 4) ( c and d ). e-h NRVMs were infected with Ad-USP18 and exposed to H/R. Scale bar=26 μm (top) and 50 μm (bottom). Representative confocal microscopy images showing mitochondrial morphology probed with MitoTracker Red ( n= 6) and quantification of mitochondrial length, mitochondrial fragmentation, and mitochondrial DNA levels ( n= 6) ( e and f ). Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in NRVMs in the indicated groups ( n= 4) ( g and h ). ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001, ns non-significant. US P 18. Ubiquitin-specific protease 18; H/R. Hypoxia/reoxygenation; NC. Negative control; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; NRVMs. Neonatal rat ventricular cardiomyocytes; ATP. Adenosine triphosphate; OCR. Oxygen consumption rate.

Techniques: Transfection, Confocal Microscopy, Infection, Ubiquitin Proteomics, Negative Control

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.

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.

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

$Parkin knockdown counteracts the protection of USP18 deficiency in vivo. USP18-cKO mice were infected with AAV9-shParkin and subjected to I/R surgery. a Parkin protein levels in mouse hearts infected with AAV9-shParkin ( n= 4). b TTC staining of heart tissue 24 h post-I/R in each group ( n= 5). Scale bar=0.5 cm. c Serum levels of cTnI, CK-MB, and LDH in AAV9-shParkin-infected mice 4 h after I/R surgery ( n= 5). d DNA fragmentation and cleaved caspase-3 activity in heart tissue from AAV9-shParkin-infected mice 24 h after I/R injury ( n= 6). e Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in mitochondria in the indicated groups ( n= 4). f Mitochondrial DNA ( n= 6) and complexes I and II–III activity ( n= 5) 24 h post-I/R in each group. g Representative electron microscopy images of heart sections 24 h post-I/R. The mitochondrial volume density and percent of mitochondria with cristae loss were measured in each group ( n= 5). Scale bar=10 μm (top) and 6 μm (bottom). h Protein levels of P62, ubiquitinated proteins (Ub), and LC3II in mitochondria from heart tissue 24 h after I/R injury ( n= 4). i H&E staining and quantitative statistical analysis of cell size ( n= 5) in USP18-cKO mice and AAV9-shParkin-infected mice 4 weeks after I/R injury. Scale bar=1 μm (top) and 50 μm (bottom). j Heart weight-to-tibia length ratio (HW/TL) ( n= 6) in each group. k Representative B-mode and M-mode echocardiographic images of the left ventricle from USP18-cKO mice and AAV9-shParkin-infected mice 4 weeks after I/R injury. l Cardiac function of USP18-cKO mice and AAV9-shParkin-infected mice 4 weeks after I/R injury ( n= 6). ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; cTnI. Cardiac Troponin I; CK-MB. Creatine kinase-MB isoenzyme; LDH. Lactate dehydrogenase; NC. Negative control; AAV9. Adeno-associated virus serotype 9; AAV9-USP18. Adeno-associated virus serotype 9 encoding USP18; TTC. 2,3,5-triphenyltetrazolium chloride; ATP. Adenosine triphosphate; H&E. Hematoxylin and eosin; PSR. Picrosirius red; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; LVIDd. Left ventricle internal diameter at diastole; LVISd. Left ventricle internal diameter at systole; LVEF. Left ventricle ejection fraction; LVFS. Left ventricle fractional shortening; HR. Heart rate.$

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: Parkin knockdown counteracts the protection of USP18 deficiency in vivo. USP18-cKO mice were infected with AAV9-shParkin and subjected to I/R surgery. a Parkin protein levels in mouse hearts infected with AAV9-shParkin ( n= 4). b TTC staining of heart tissue 24 h post-I/R in each group ( n= 5). Scale bar=0.5 cm. c Serum levels of cTnI, CK-MB, and LDH in AAV9-shParkin-infected mice 4 h after I/R surgery ( n= 5). d DNA fragmentation and cleaved caspase-3 activity in heart tissue from AAV9-shParkin-infected mice 24 h after I/R injury ( n= 6). e Oxygen consumption rate (OCR) and quantitative statistical analysis of basal respiration, ATP-related respiration, maximal respiration, and spare respiratory capacity in mitochondria in the indicated groups ( n= 4). f Mitochondrial DNA ( n= 6) and complexes I and II–III activity ( n= 5) 24 h post-I/R in each group. g Representative electron microscopy images of heart sections 24 h post-I/R. The mitochondrial volume density and percent of mitochondria with cristae loss were measured in each group ( n= 5). Scale bar=10 μm (top) and 6 μm (bottom). h Protein levels of P62, ubiquitinated proteins (Ub), and LC3II in mitochondria from heart tissue 24 h after I/R injury ( n= 4). i H&E staining and quantitative statistical analysis of cell size ( n= 5) in USP18-cKO mice and AAV9-shParkin-infected mice 4 weeks after I/R injury. Scale bar=1 μm (top) and 50 μm (bottom). j Heart weight-to-tibia length ratio (HW/TL) ( n= 6) in each group. k Representative B-mode and M-mode echocardiographic images of the left ventricle from USP18-cKO mice and AAV9-shParkin-infected mice 4 weeks after I/R injury. l Cardiac function of USP18-cKO mice and AAV9-shParkin-infected mice 4 weeks after I/R injury ( n= 6). ⁎ P <0.05, ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; cTnI. Cardiac Troponin I; CK-MB. Creatine kinase-MB isoenzyme; LDH. Lactate dehydrogenase; NC. Negative control; AAV9. Adeno-associated virus serotype 9; AAV9-USP18. Adeno-associated virus serotype 9 encoding USP18; TTC. 2,3,5-triphenyltetrazolium chloride; ATP. Adenosine triphosphate; H&E. Hematoxylin and eosin; PSR. Picrosirius red; FCCP. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone; LVIDd. Left ventricle internal diameter at diastole; LVISd. Left ventricle internal diameter at systole; LVEF. Left ventricle ejection fraction; LVFS. Left ventricle fractional shortening; HR. Heart rate.

Techniques: Knockdown, In Vivo, Infection, Staining, Activity Assay, Electron Microscopy, Ubiquitin Proteomics, Negative Control, Virus

USP18 inhibits mitophagy degradation and facilitates cardiac I/R injury through deubiquitinating and upregulating PTEN-L. a The protein levels of PTEN-L and PTEN in USP18-cKO mouse hearts 24 h after I/R injury ( n= 4). ⁎⁎⁎⁎ P <0.0001, ns non-significant. b The protein levels of PTEN-L and PTEN in AAV9-USP18-infected mouse hearts 24 h after I/R injury ( n= 4). ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001, ns non-significant. c Co-IP of USP18 and PTEN-L in NRVMs (left); NRVMs were transfected with HA-PTEN-L and Flag-USP18 (middle); Co-IP of Flag-USP18 and HA-PTEN-L in NRVMs (right). d Endogenous Co-IP of USP18 and PTEN-L in NRVMs subjected to H/R injury. e Endogenous Co-IP of USP18 and PTEN-L in hearts subjected to I/R injury. f PTEN-L ubiquitination (Ub) levels assessed by CO-IP in NRVMs subjected to H/R injury (top) and hearts subjected to I/R injury (bottom). g NRVMs were transfected with Ad-USP18 or USP18 siRNA or HA-PTEN-L and Myc-Ub and treated with MG132. Co-IP of Myc-Ub and HA-PTEN-L. h Schematic representations of t h e domains of PTEN-L involved in binding to USP18. Full-length PTEN-L or truncated PTEN-L was coexpressed with USP18 in HEK293T cells. Cells were subjected to immunoprecipitation with an anti-Myc antibody or an anti-HA antibody, followed by immunoblotting with the indicated antibodies. i Schematic representations of USP18 residues involved in binding to PTEN-L. Full-length USP18 or USP18 truncations were coexpressed with PTEN-L in HEK293T cells. Cells were subjected to immunoprecipitation with an anti-Myc antibody or an anti-HA antibody, followed by immunoblotting with the indicated antibodies. j Ub assay of PTEN-L in HEK293T cells cotransfected with Myc-USP18, Myc-USP18-mut, and Flag-PTEN-L and treated with 10 μmol/L MG132. k, l NRVMs were transfected with scRNA or USP18 siRNA ( k ), infected with Ad-NC or Ad-USP18 ( l ), and then treated with cycloheximide (CHX, 10 μmol/L) for the indicated time periods. Representative immunoblot analysis of PTEN-L protein levels in each group. ⁎⁎⁎⁎ P <0.0001 vs . scRNA or Ad-NC. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; H/R. Hypoxia-reoxygenation; PTEN. Phosphatase and tensin homolog; PTEN-L. Phosphatase and tensin homolog-long; AAV9. Adeno-associated virus serotype 9; NC. Negative control.

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 inhibits mitophagy degradation and facilitates cardiac I/R injury through deubiquitinating and upregulating PTEN-L. a The protein levels of PTEN-L and PTEN in USP18-cKO mouse hearts 24 h after I/R injury ( n= 4). ⁎⁎⁎⁎ P <0.0001, ns non-significant. b The protein levels of PTEN-L and PTEN in AAV9-USP18-infected mouse hearts 24 h after I/R injury ( n= 4). ⁎⁎⁎ P <0.001, ⁎⁎⁎⁎ P <0.0001, ns non-significant. c Co-IP of USP18 and PTEN-L in NRVMs (left); NRVMs were transfected with HA-PTEN-L and Flag-USP18 (middle); Co-IP of Flag-USP18 and HA-PTEN-L in NRVMs (right). d Endogenous Co-IP of USP18 and PTEN-L in NRVMs subjected to H/R injury. e Endogenous Co-IP of USP18 and PTEN-L in hearts subjected to I/R injury. f PTEN-L ubiquitination (Ub) levels assessed by CO-IP in NRVMs subjected to H/R injury (top) and hearts subjected to I/R injury (bottom). g NRVMs were transfected with Ad-USP18 or USP18 siRNA or HA-PTEN-L and Myc-Ub and treated with MG132. Co-IP of Myc-Ub and HA-PTEN-L. h Schematic representations of t h e domains of PTEN-L involved in binding to USP18. Full-length PTEN-L or truncated PTEN-L was coexpressed with USP18 in HEK293T cells. Cells were subjected to immunoprecipitation with an anti-Myc antibody or an anti-HA antibody, followed by immunoblotting with the indicated antibodies. i Schematic representations of USP18 residues involved in binding to PTEN-L. Full-length USP18 or USP18 truncations were coexpressed with PTEN-L in HEK293T cells. Cells were subjected to immunoprecipitation with an anti-Myc antibody or an anti-HA antibody, followed by immunoblotting with the indicated antibodies. j Ub assay of PTEN-L in HEK293T cells cotransfected with Myc-USP18, Myc-USP18-mut, and Flag-PTEN-L and treated with 10 μmol/L MG132. k, l NRVMs were transfected with scRNA or USP18 siRNA ( k ), infected with Ad-NC or Ad-USP18 ( l ), and then treated with cycloheximide (CHX, 10 μmol/L) for the indicated time periods. Representative immunoblot analysis of PTEN-L protein levels in each group. ⁎⁎⁎⁎ P <0.0001 vs . scRNA or Ad-NC. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; H/R. Hypoxia-reoxygenation; PTEN. Phosphatase and tensin homolog; PTEN-L. Phosphatase and tensin homolog-long; AAV9. Adeno-associated virus serotype 9; NC. Negative control.

Techniques: Infection, Co-Immunoprecipitation Assay, Transfection, Ubiquitin Proteomics, Binding Assay, Immunoprecipitation, Western Blot, Virus, Negative Control

Summary of the key findings in this paper. Cardiac I/R injury upregulates USP18 in both mouse and human hearts. USP18 interacts with PTEN-L, inhibiting Parkin phosphorylation and mitochondrial translocation, thereby suppressing mitophagy and exacerbating mitochondrial dysfunction and myocardial injury. PTEN-L additionally promotes cardiac damage via a paracrine mechanism. Genetic or pharmacological targeting of the USP18-PTEN-L axis restores mitophagy and protects against I/R-induced cardiac injury, representing a potential therapeutic strategy. I/R. Ischemia/reperfusion; PTEN-L. Phosphatase and tensin homolog-long; USP18. Ubiquitin-specific protease 18; Ub. Ubiquitination.

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: Summary of the key findings in this paper. Cardiac I/R injury upregulates USP18 in both mouse and human hearts. USP18 interacts with PTEN-L, inhibiting Parkin phosphorylation and mitochondrial translocation, thereby suppressing mitophagy and exacerbating mitochondrial dysfunction and myocardial injury. PTEN-L additionally promotes cardiac damage via a paracrine mechanism. Genetic or pharmacological targeting of the USP18-PTEN-L axis restores mitophagy and protects against I/R-induced cardiac injury, representing a potential therapeutic strategy. I/R. Ischemia/reperfusion; PTEN-L. Phosphatase and tensin homolog-long; USP18. Ubiquitin-specific protease 18; Ub. Ubiquitination.

Techniques: Phospho-proteomics, Translocation Assay, Ubiquitin Proteomics

Review

cytochrome c oxidase subunit 4 (Cell Signaling Technology Inc)

Structured Review

Images

Related Articles

other:

Immunohistochemistry:

Paraffin Section:

Staining:

Incubation:

Western Blot:

Similar Products

Image Search Results