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Elabscience Biotechnology myeloperoxidase activity assay
Myeloperoxidase Activity Assay, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 180 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mpo/Myeloperoxidase+(MPO)+Activity+Assay+Kit/10__3390_slash_md24100361-268-6-9
Average 96 stars, based on 180 article reviews

myeloperoxidase activity assay - by Bioz Stars, 2026-11

96/100 stars

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Article Title: hUC-MSCs exosomal miR-451 alleviated acute lung injury by modulating macrophage M2 polarization via regulating MIF-PI3K-AKT signaling pathway.
Article Snippet: Funding information Key scientific research project of Anhui Provincial Health Commission in 2021, Grant/Award Number: ahwj2021a042; The tenth batch of "3221" industrial innovation and scientific research projects in Bengbu City Abstract In the previous study, we have proved that exosomal miR-451 from human umbilical cord mesenchymal stem cells (hUC-MSCs) attenuated burn-induced acute lung injury (ALI).. However, the mechanism of exosomal miR-451 in ALI remains unclear.. Therefore, this study aimed to study the molecular mechanism of hUC-MSCs-derived exosomal miR-451 on ALI by regulating macrophage polarization.

Article Title: T-cell apoptosis is more susceptible to overactivated neutrophils delivered-NETs-MPO/NE complex.
Article Snippet: Sepsis, characterized by infection-induced systemic inflammatory response, involves polymorphonuclear leukocytes (PMNs or Neutrophils)-T cell dysregulation.. However, the precise mechanisms of PMN-driven T cell apoptosis via neutrophil extracellular traps (NETs) and their protease cargo (myeloperoxidase [MPO], neutrophil elastase [NE]), along with potential mitochondrial transfer-mediated protection, remain unclear.. Using PMAactivated PMNs in coculture systems, we assessed NETs formation, T cell apoptosis/proliferation/exhaustion, DNA damage, and mitochondrial function through flow cytometry, electron microscopy, immunofluorescence, and molecular assays, with validation in LPS-challenged mice.

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Article Title: Multi-omics analysis reveals the pre-protective mechanism of Dendrobium flexicaule polysaccharide against alcohol-induced gastric mucosal injury.
Article Snippet: Dendrobium flexicaule (DF) is an endemic plant primarily found in the mountains of central China with important medicinal and edible values.. In traditional Chinese medicine, DF has the effects of nourishing stomach and “Yin”, and clearing heat.. At present, no studies have explored the mechanisms by which Dendrobium flexicaule polysaccharides (DFP) exert pre-protect effects against alcohol-induced gastric mucosal injury.

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Identification of a cross-reactive monoclonal antibody against various serotypes of Salmonella . (A) Enzyme-linked immunosorbent assay was used to screen hybridomas for reactivity to a range of heat-killed bacteria, including five Salmonella serotypes. Approximately 2000 clones were tested, and representative data from selected clone L2-1H11 are shown. (B) Reactivity of purified L2-1H11 monoclonal antibody to the indicated Salmonella serotypes, as determined by flow cytometry. (C) Stimulated emission depletion microscopy images and intensity (regions indicated by white line) of Salmonella Typhimurium stained with L2-1H11 monoclonal antibody and FM4-64, which labels the bacterial cell surface.

Journal: Frontiers in Immunology

Article Title: Oligopeptide-binding protein A is a potential target for combating Salmonella infection

doi: 10.3389/fimmu.2026.1943609

Figure Lengend Snippet: Identification of a cross-reactive monoclonal antibody against various serotypes of Salmonella . (A) Enzyme-linked immunosorbent assay was used to screen hybridomas for reactivity to a range of heat-killed bacteria, including five Salmonella serotypes. Approximately 2000 clones were tested, and representative data from selected clone L2-1H11 are shown. (B) Reactivity of purified L2-1H11 monoclonal antibody to the indicated Salmonella serotypes, as determined by flow cytometry. (C) Stimulated emission depletion microscopy images and intensity (regions indicated by white line) of Salmonella Typhimurium stained with L2-1H11 monoclonal antibody and FM4-64, which labels the bacterial cell surface.

Article Snippet: Myeloperoxidase levels in the supernatant were measured using a myeloperoxidase ELISA kit (Hycult Biotech Inc.).

Techniques: Enzyme-linked Immunosorbent Assay, Bacteria, Clone Assay, Purification, Flow Cytometry, Microscopy, Staining

L2-1H11 antibody protects against Salmonella infection. (A) Salmonella Typhimurium cells were treated with 100 μg of isotype control or L2-1H11 antibody, incubated at 37°C for 1 h, and used to infect HT29 cells. Cell death was quantified on the basis of the amount of lactate dehydrogenase released into the culture supernatant (n = 6). OD, optical density. (B, C) After adhesion and invasion of HT29 cells by Salmonella , the HT29 cells were washed, lysed, and the lysates were plated on Salmonella -Shigella agar and cultured overnight. The numbers of colony-forming units (CFUs) were counted (n = 6). (D) Salmonella Typhimurium (1.0 × 106 CFU) was treated with 100 μg of isotype control or L2-1H11 antibody and incubated at 37°C for 1 h. Mice were orally infected with the treated bacteria using a gavage tube. Fecal samples were collected daily, suspended in LB broth, plated on Salmonella-Shigella agar, incubated overnight, and the number of CFUs was counted. (E) The intestines were excised 2 and 4 days post-infection with Salmonella Typhimurium conjugated with L2-1H11 antibody or anti- Salmonella Typhimurium LPS antibody, and neutrophils in the lamina propria were counted on the basis of flow cytometric analysis data. (F) Immunohistological analysis of the ileum 4 days post-infection. Neutrophils, epithelial cells, and nuclei were stained with Ly6G monoclonal antibody (red), EpCAM monoclonal antibody (green), and DAPI (blue), respectively. (G) Fecal samples from infected mice were collected daily, and the concentration of myeloperoxidase (MPO), which is specifically secreted by neutrophils, was quantified by enzyme-linked immunosorbent assay. (H) Salmonella Typhimurium cells were treated with 1 μg of isotype control or L2-1H11 antibody and incubated at 37°C for 1 h. They were then stained by using a Live/Dead BacLight Bacterial Viability Kit, and the ratio of live to dead Salmonella was determined by using a flow cytometer. (I) Salmonella Typhimurium cells were treated with 100 μg of isotype control, L2-1H11 antibody, or anti- Salmonella Typhimurium LPS antibody and incubated at 37°C for 1 h, transferred to LB broth or M9 minimal medium, and bacterial growth was assessed at 0, 1, 2, 4, 6, 8, 16, and 24 h. Data are representative of two independent experiments and are presented as mean ± SD. Two-tailed Student’s t-test (A–C) , one-way ANOVA (E) , two-way ANOVA (I) and repeated-measures two-way ANOVA (D, G) : *P < 0.05, ***P < 0.001. Scale bars, 100 μm.

Journal: Frontiers in Immunology

Article Title: Oligopeptide-binding protein A is a potential target for combating Salmonella infection

doi: 10.3389/fimmu.2026.1943609

Figure Lengend Snippet: L2-1H11 antibody protects against Salmonella infection. (A) Salmonella Typhimurium cells were treated with 100 μg of isotype control or L2-1H11 antibody, incubated at 37°C for 1 h, and used to infect HT29 cells. Cell death was quantified on the basis of the amount of lactate dehydrogenase released into the culture supernatant (n = 6). OD, optical density. (B, C) After adhesion and invasion of HT29 cells by Salmonella , the HT29 cells were washed, lysed, and the lysates were plated on Salmonella -Shigella agar and cultured overnight. The numbers of colony-forming units (CFUs) were counted (n = 6). (D) Salmonella Typhimurium (1.0 × 106 CFU) was treated with 100 μg of isotype control or L2-1H11 antibody and incubated at 37°C for 1 h. Mice were orally infected with the treated bacteria using a gavage tube. Fecal samples were collected daily, suspended in LB broth, plated on Salmonella-Shigella agar, incubated overnight, and the number of CFUs was counted. (E) The intestines were excised 2 and 4 days post-infection with Salmonella Typhimurium conjugated with L2-1H11 antibody or anti- Salmonella Typhimurium LPS antibody, and neutrophils in the lamina propria were counted on the basis of flow cytometric analysis data. (F) Immunohistological analysis of the ileum 4 days post-infection. Neutrophils, epithelial cells, and nuclei were stained with Ly6G monoclonal antibody (red), EpCAM monoclonal antibody (green), and DAPI (blue), respectively. (G) Fecal samples from infected mice were collected daily, and the concentration of myeloperoxidase (MPO), which is specifically secreted by neutrophils, was quantified by enzyme-linked immunosorbent assay. (H) Salmonella Typhimurium cells were treated with 1 μg of isotype control or L2-1H11 antibody and incubated at 37°C for 1 h. They were then stained by using a Live/Dead BacLight Bacterial Viability Kit, and the ratio of live to dead Salmonella was determined by using a flow cytometer. (I) Salmonella Typhimurium cells were treated with 100 μg of isotype control, L2-1H11 antibody, or anti- Salmonella Typhimurium LPS antibody and incubated at 37°C for 1 h, transferred to LB broth or M9 minimal medium, and bacterial growth was assessed at 0, 1, 2, 4, 6, 8, 16, and 24 h. Data are representative of two independent experiments and are presented as mean ± SD. Two-tailed Student’s t-test (A–C) , one-way ANOVA (E) , two-way ANOVA (I) and repeated-measures two-way ANOVA (D, G) : *P < 0.05, ***P < 0.001. Scale bars, 100 μm.

Article Snippet: Myeloperoxidase levels in the supernatant were measured using a myeloperoxidase ELISA kit (Hycult Biotech Inc.).

Techniques: Infection, Control, Incubation, Cell Culture, Bacteria, Fecal/Stool Samples, Analysis, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Two Tailed Test

L2-1H11 antibody recognizes oligopeptide-binding protein A (OppA). Immunoprecipitation (A) and western blot analysis (B) of a lysate of Salmonella cultured in LB broth in the presence of L2-1H11 antibody. (C) NuPAGE electrophoresis of recombinant OppA protein and western blot analysis using L2-1H11. (D) Binding affinity of L2-1H11 to OppA was quantitatively analyzed by surface plasmon resonance assay. (E) Comparison of amino acids 290–302 of OppA from the indicated Salmonella serotypes and Escherichia coli DH5α. (F) L2-1H11 antibody binding to recombinant OppA derived from Salmonella Typhimurium or E. coli was evaluated by enzyme-linked immunosorbent assay to assess potential off-target cross-reactivity with commensal bacteria. Absorbance was measured at 450 nm. Data are shown as mean ± SD of two independent experiments. (G) Three-dimensional structure of Salmonella Typhimurium OppA shown alongside its single-point mutant (R291H), as predicted using the AlphaFold structure prediction tool. The location of the substituted amino acid is indicated by red circles.

Journal: Frontiers in Immunology

Article Title: Oligopeptide-binding protein A is a potential target for combating Salmonella infection

doi: 10.3389/fimmu.2026.1943609

Figure Lengend Snippet: L2-1H11 antibody recognizes oligopeptide-binding protein A (OppA). Immunoprecipitation (A) and western blot analysis (B) of a lysate of Salmonella cultured in LB broth in the presence of L2-1H11 antibody. (C) NuPAGE electrophoresis of recombinant OppA protein and western blot analysis using L2-1H11. (D) Binding affinity of L2-1H11 to OppA was quantitatively analyzed by surface plasmon resonance assay. (E) Comparison of amino acids 290–302 of OppA from the indicated Salmonella serotypes and Escherichia coli DH5α. (F) L2-1H11 antibody binding to recombinant OppA derived from Salmonella Typhimurium or E. coli was evaluated by enzyme-linked immunosorbent assay to assess potential off-target cross-reactivity with commensal bacteria. Absorbance was measured at 450 nm. Data are shown as mean ± SD of two independent experiments. (G) Three-dimensional structure of Salmonella Typhimurium OppA shown alongside its single-point mutant (R291H), as predicted using the AlphaFold structure prediction tool. The location of the substituted amino acid is indicated by red circles.

Article Snippet: Myeloperoxidase levels in the supernatant were measured using a myeloperoxidase ELISA kit (Hycult Biotech Inc.).

Techniques: Binding Assay, Immunoprecipitation, Western Blot, Analysis, Cell Culture, Electrophoresis, Recombinant, SPR Assay, Comparison, Derivative Assay, Enzyme-linked Immunosorbent Assay, Bacteria, Mutagenesis

Protective effects and immune responses induced by oral immunization with OppA in mice against salmonellosis. Mice were orally immunized with either phosphate-buffered saline (mock) or OppA once a week for four times. (A) One week after the final immunization, a feces sample was collected for the detection of Salmonella -specific fecal IgA by enzyme-linked immunosorbent assay (n = 10). (B, C) ECAR and OCR were measured in Salmonella treated with the fecal suspension collected from mice immunized orally with mock or OppA (n = 4). (D) Number of Salmonella in feces from immunized with mock or OppA and infected mice was monitored daily. Fecal samples were suspended in LB broth, and the supernatant of the fecal slurry was plated on Salmonella -Shigella agar and cultured overnight to count colony-forming units (CFUs). (E) Myeloperoxidase (MPO) levels were quantified daily in feces from immunized and infected mice by enzyme-linked immunosorbent assay (n = 4). (F) Four days post-infection, the intestines were collected from mice orally immunized with mock or OppA, and the number of neutrophils in the lamina propria was quantified by flow cytometric analysis. (G) Survival rates of mice orally immunized with mock or OppA were monitored for 2 weeks post-infection with Salmonella . (H) CFUs of Salmonella in the liver and spleen were assessed on day 4 post-infection in mice that had been orally immunized and then infected with Salmonella . (I) Serum samples were collected from naïve mice (non-immunization) and mice orally immunized with mock or OppA on day 4 post-infection, and activity of alanine transaminase (ALT) and aspartate aminotransferase (AST) and (J) concentrations of cytokines or a chemokine were determined (n = 4). IL, interleukin; MCP, monocyte chemoattractant protein; TNF, tumor necrosis factor. Data are representative of two independent experiments and are presented as mean ± SD. Two-tailed Student’s t-test (A–G, J) , one-way ANOVA (H, I) , repeated-measures two-way ANOVA (D, E) , and log-rank (Mantel-Cox) test (G) : **P < 0.01, ***P < 0.001.

Journal: Frontiers in Immunology

Article Title: Oligopeptide-binding protein A is a potential target for combating Salmonella infection

doi: 10.3389/fimmu.2026.1943609

Figure Lengend Snippet: Protective effects and immune responses induced by oral immunization with OppA in mice against salmonellosis. Mice were orally immunized with either phosphate-buffered saline (mock) or OppA once a week for four times. (A) One week after the final immunization, a feces sample was collected for the detection of Salmonella -specific fecal IgA by enzyme-linked immunosorbent assay (n = 10). (B, C) ECAR and OCR were measured in Salmonella treated with the fecal suspension collected from mice immunized orally with mock or OppA (n = 4). (D) Number of Salmonella in feces from immunized with mock or OppA and infected mice was monitored daily. Fecal samples were suspended in LB broth, and the supernatant of the fecal slurry was plated on Salmonella -Shigella agar and cultured overnight to count colony-forming units (CFUs). (E) Myeloperoxidase (MPO) levels were quantified daily in feces from immunized and infected mice by enzyme-linked immunosorbent assay (n = 4). (F) Four days post-infection, the intestines were collected from mice orally immunized with mock or OppA, and the number of neutrophils in the lamina propria was quantified by flow cytometric analysis. (G) Survival rates of mice orally immunized with mock or OppA were monitored for 2 weeks post-infection with Salmonella . (H) CFUs of Salmonella in the liver and spleen were assessed on day 4 post-infection in mice that had been orally immunized and then infected with Salmonella . (I) Serum samples were collected from naïve mice (non-immunization) and mice orally immunized with mock or OppA on day 4 post-infection, and activity of alanine transaminase (ALT) and aspartate aminotransferase (AST) and (J) concentrations of cytokines or a chemokine were determined (n = 4). IL, interleukin; MCP, monocyte chemoattractant protein; TNF, tumor necrosis factor. Data are representative of two independent experiments and are presented as mean ± SD. Two-tailed Student’s t-test (A–G, J) , one-way ANOVA (H, I) , repeated-measures two-way ANOVA (D, E) , and log-rank (Mantel-Cox) test (G) : **P < 0.01, ***P < 0.001.

Article Snippet: Myeloperoxidase levels in the supernatant were measured using a myeloperoxidase ELISA kit (Hycult Biotech Inc.).

Techniques: Saline, Enzyme-linked Immunosorbent Assay, Suspension, Infection, Fecal/Stool Samples, Cell Culture, Analysis, Blood/Plasma/Serum Samples, Activity Assay, Two Tailed Test

NOD2 deficiency ameliorates cigarette smoke-induced lung inflammation and injury in vivo The functional role of NOD2 was evaluated in a cigarette smoke (CS)-induced mouse model of COPD using wild-type (wt) and NOD2-knockout (NOD2-/-) mice, with lentiviral-mediated NOD2 re-expression (NOD2-OE) in NOD2-/- COPD mice. ( A ) Schematic of the in vivo experimental design. ( B ) qRT-PCR analysis of NOD2 mRNA expression in lung tissue. ( C ) Representative images of hematoxylin and eosin (H&E) staining of lung tissue sections. Scale bars: 200 μm in the upper panels and 100 μm in the enlarged lower panels. ( D ) Representative images of Masson’s trichrome staining for collagen deposition in lung tissue sections. Scale bar: 100 μm. ( E ) Quantification of total cells, neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF), and ELISA measurements of TNF-α, IL-6, and CXCL1 levels in BALF supernatant. ( F ) Representative images of IHC staining for E-cadherin and N-cadherin in lung tissue. Scale bar: 50 μm. ( G ) Quantification of the average optical density (AOD) for E-cadherin and N-cadherin IHC staining. ( H ) qRT-PCR analysis of senescence-associated secretory phenotype (SASP) factors (CSF3, CCL3, and IL1α) in lung tissue. Data are presented as mean ± SD. Data were analyzed using one-way ANOVA followed by Tukey’s test. * p < 0.05 and ** p < 0.01 Control vs. wt-COPD groups; # p < 0.05 and ## p < 0.01 wt-COPD vs. NOD2 −/− -COPD groups; & p < 0.05 and && p < 0.01 NOD2 −/− -COPD + vector vs. NOD2 −/− -COPD + NOD2-OE groups

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: NOD2 deficiency ameliorates cigarette smoke-induced lung inflammation and injury in vivo The functional role of NOD2 was evaluated in a cigarette smoke (CS)-induced mouse model of COPD using wild-type (wt) and NOD2-knockout (NOD2-/-) mice, with lentiviral-mediated NOD2 re-expression (NOD2-OE) in NOD2-/- COPD mice. ( A ) Schematic of the in vivo experimental design. ( B ) qRT-PCR analysis of NOD2 mRNA expression in lung tissue. ( C ) Representative images of hematoxylin and eosin (H&E) staining of lung tissue sections. Scale bars: 200 μm in the upper panels and 100 μm in the enlarged lower panels. ( D ) Representative images of Masson’s trichrome staining for collagen deposition in lung tissue sections. Scale bar: 100 μm. ( E ) Quantification of total cells, neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF), and ELISA measurements of TNF-α, IL-6, and CXCL1 levels in BALF supernatant. ( F ) Representative images of IHC staining for E-cadherin and N-cadherin in lung tissue. Scale bar: 50 μm. ( G ) Quantification of the average optical density (AOD) for E-cadherin and N-cadherin IHC staining. ( H ) qRT-PCR analysis of senescence-associated secretory phenotype (SASP) factors (CSF3, CCL3, and IL1α) in lung tissue. Data are presented as mean ± SD. Data were analyzed using one-way ANOVA followed by Tukey’s test. * p < 0.05 and ** p < 0.01 Control vs. wt-COPD groups; # p < 0.05 and ## p < 0.01 wt-COPD vs. NOD2 −/− -COPD groups; & p < 0.05 and && p < 0.01 NOD2 −/− -COPD + vector vs. NOD2 −/− -COPD + NOD2-OE groups

Article Snippet: Additionally, the commercial ELISA kits (Human and mouse MPO ELISA Kits, Cat. # CSB-E08721h, CSB-E08723m, CUSABIO, Wuhan, China; Human and mouse neutrophil elastase ELISA Kit, Cat. # CSB-EL007587HU, CSB-E04804m, CUSABIO) were applied to quantify the levels of MPO and neutrophil elastase released into cell culture supernatants as per the instructions of the manufacturer.

Techniques: In Vivo, Functional Assay, Knock-Out, Expressing, Quantitative RT-PCR, Analysis, Tissue, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Control, Plasmid Preparation

NOD2 promotes Neutrophil Extracellular Trap (NET) formation in the CS-induced COPD mouse model The association between NOD2, autophagy, and Neutrophil Extracellular Trap (NET) formation was investigated in the lung tissue of mice from the CS-induced COPD model. ( A) Representative immunofluorescence images showing co-localization of the neutrophil marker Ly6G (green) and the autophagy marker LC3B (red), with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( B ) Representative immunofluorescence images showing staining for the NET markers citrullinated histone H3 (CitH3, green) and myeloperoxidase (MPO, red), with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( C ) Quantification of the NET biomarkers CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. ( D ) Immunoblotting analysis of the neutrophil marker Ly6G and the NET marker CitH3 in whole-lung tissue lysates. ( E ) Representative immunofluorescence images showing colocalization of the neutrophil marker Ly6G (red), the NET marker CitH3 (green), and NOD2 (pink) in lung tissue, with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. Data are presented as mean ± SD. C and D were analyzed using one-way ANOVA followed by Tukey’s test. ** p < 0.01 Control vs. wt -COPD groups; and ## p < 0.01 wt -COPD vs. NOD2 −/− -COPD groups; && p < 0.01 NOD2 −/− -COPD + vector vs. NOD2 −/− -COPD + NOD2-OE groups

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: NOD2 promotes Neutrophil Extracellular Trap (NET) formation in the CS-induced COPD mouse model The association between NOD2, autophagy, and Neutrophil Extracellular Trap (NET) formation was investigated in the lung tissue of mice from the CS-induced COPD model. ( A) Representative immunofluorescence images showing co-localization of the neutrophil marker Ly6G (green) and the autophagy marker LC3B (red), with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( B ) Representative immunofluorescence images showing staining for the NET markers citrullinated histone H3 (CitH3, green) and myeloperoxidase (MPO, red), with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( C ) Quantification of the NET biomarkers CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. ( D ) Immunoblotting analysis of the neutrophil marker Ly6G and the NET marker CitH3 in whole-lung tissue lysates. ( E ) Representative immunofluorescence images showing colocalization of the neutrophil marker Ly6G (red), the NET marker CitH3 (green), and NOD2 (pink) in lung tissue, with nuclei counterstained with DAPI (blue). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. Data are presented as mean ± SD. C and D were analyzed using one-way ANOVA followed by Tukey’s test. ** p < 0.01 Control vs. wt -COPD groups; and ## p < 0.01 wt -COPD vs. NOD2 −/− -COPD groups; && p < 0.01 NOD2 −/− -COPD + vector vs. NOD2 −/− -COPD + NOD2-OE groups

Techniques: Tissue, Immunofluorescence, Marker, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Analysis, Control, Plasmid Preparation

Neutrophil depletion minimizes genotype-dependent differences in CS-induced lung injury, inflammation, fibrosis, and NET formation The dependence of NOD2-associated COPD pathology on neutrophils was evaluated by depleting neutrophils in CS-exposed wild-type (wt) and NOD2 −/− mice. Mice were randomly assigned to the wt -COPD + IgG, wt -COPD + Anti-Ly6G, NOD2 −/− -COPD + IgG, and NOD2 −/− -COPD + Anti-Ly6G groups ( n = 6 per group). ( A ) Schematic of the experimental design. Mice were exposed to CS for 12 weeks and received 200 µg of anti-Ly6G antibody or an equivalent dose of isotype IgG intraperitoneally three times per week from week 2 until the end of the experiment. ( B ) Representative flow-cytometry plots and quantification of CD11b + Ly6G + neutrophils in lung tissue and peripheral blood. ( C ) Representative H&E-stained images of lung tissue sections. Scale bars: 200 μm in the upper panels and 100 μm in the enlarged lower panels. ( D ) Representative images of Masson’s trichrome staining for pulmonary collagen deposition. Scale bars: 100 μm. ( E ) Quantification of total cells, neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF), together with ELISA measurements of TNF-α, IL-6, and CXCL1 levels in BALF supernatants. ( F ) Quantification of CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. Data are presented as mean ± SD. Data were analyzed using two-way ANOVA followed by Tukey’s test. ** p < 0.01 vs. the wt -COPD + IgG group; ## p < 0.01 vs. the wt-COPD + Anti-Ly6G group; & p < 0.05, & p < 0.01 vs. the NOD2 −/− -COPD + IgG group

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: Neutrophil depletion minimizes genotype-dependent differences in CS-induced lung injury, inflammation, fibrosis, and NET formation The dependence of NOD2-associated COPD pathology on neutrophils was evaluated by depleting neutrophils in CS-exposed wild-type (wt) and NOD2 −/− mice. Mice were randomly assigned to the wt -COPD + IgG, wt -COPD + Anti-Ly6G, NOD2 −/− -COPD + IgG, and NOD2 −/− -COPD + Anti-Ly6G groups ( n = 6 per group). ( A ) Schematic of the experimental design. Mice were exposed to CS for 12 weeks and received 200 µg of anti-Ly6G antibody or an equivalent dose of isotype IgG intraperitoneally three times per week from week 2 until the end of the experiment. ( B ) Representative flow-cytometry plots and quantification of CD11b + Ly6G + neutrophils in lung tissue and peripheral blood. ( C ) Representative H&E-stained images of lung tissue sections. Scale bars: 200 μm in the upper panels and 100 μm in the enlarged lower panels. ( D ) Representative images of Masson’s trichrome staining for pulmonary collagen deposition. Scale bars: 100 μm. ( E ) Quantification of total cells, neutrophils, and macrophages in bronchoalveolar lavage fluid (BALF), together with ELISA measurements of TNF-α, IL-6, and CXCL1 levels in BALF supernatants. ( F ) Quantification of CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. Data are presented as mean ± SD. Data were analyzed using two-way ANOVA followed by Tukey’s test. ** p < 0.01 vs. the wt -COPD + IgG group; ## p < 0.01 vs. the wt-COPD + Anti-Ly6G group; & p < 0.05, & p < 0.01 vs. the NOD2 −/− -COPD + IgG group

Techniques: Pathology, Flow Cytometry, Tissue, Staining, Enzyme-linked Immunosorbent Assay

Knockdown of NOD2 inhibits CSE-induced NET formation, ROS production, and mitophagy in human neutrophils The role of NOD2 in cigarette smoke extract (CSE)-induced Neutrophil Extracellular Trap (NET) formation, reactive oxygen species (ROS) accumulation, and lysosome–mitochondrion spatial relationships were evaluated in differentiated HL-60-derived neutrophil-like cells. ( A ) Schematic of the in vitro experimental design and treatment groups. ( B ) Representative DAPI-stained images of extracellular DNA structures and quantification of NET formation based on DAPI-stained nuclear morphology. Scale bars: 20 μm. ( C ) Extracellular DNA levels in cell culture supernatants determined using a PicoGreen dsDNA assay. ( D ) Myeloperoxidase (MPO) levels in cell culture supernatants determined by ELISA. ( E ) Neutrophil elastase (NE) levels in cell culture supernatants determined by ELISA. ( F ) Representative flow-cytometry histograms and quantitative analysis of intracellular ROS levels measured using DCFH-DA staining. ( G ) Representative immunofluorescence images showing the spatial relationship between LAMP1-positive lysosomes (green) and TOM20-positive mitochondria (red), with nuclei counterstained with DAPI (blue). The colocation efficients for LAMP1 and TOM20 was determined by ImageJ and shown on the right. Scale bars: 20 μm. Data are presented as mean ± SD. Quantitative data were analyzed using one-way ANOVA followed by Tukey’s test. ** p < 0.01 vs. Control group; ## p < 0.01 vs. CSE + sh-NC group

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: Knockdown of NOD2 inhibits CSE-induced NET formation, ROS production, and mitophagy in human neutrophils The role of NOD2 in cigarette smoke extract (CSE)-induced Neutrophil Extracellular Trap (NET) formation, reactive oxygen species (ROS) accumulation, and lysosome–mitochondrion spatial relationships were evaluated in differentiated HL-60-derived neutrophil-like cells. ( A ) Schematic of the in vitro experimental design and treatment groups. ( B ) Representative DAPI-stained images of extracellular DNA structures and quantification of NET formation based on DAPI-stained nuclear morphology. Scale bars: 20 μm. ( C ) Extracellular DNA levels in cell culture supernatants determined using a PicoGreen dsDNA assay. ( D ) Myeloperoxidase (MPO) levels in cell culture supernatants determined by ELISA. ( E ) Neutrophil elastase (NE) levels in cell culture supernatants determined by ELISA. ( F ) Representative flow-cytometry histograms and quantitative analysis of intracellular ROS levels measured using DCFH-DA staining. ( G ) Representative immunofluorescence images showing the spatial relationship between LAMP1-positive lysosomes (green) and TOM20-positive mitochondria (red), with nuclei counterstained with DAPI (blue). The colocation efficients for LAMP1 and TOM20 was determined by ImageJ and shown on the right. Scale bars: 20 μm. Data are presented as mean ± SD. Quantitative data were analyzed using one-way ANOVA followed by Tukey’s test. ** p < 0.01 vs. Control group; ## p < 0.01 vs. CSE + sh-NC group

Techniques: Knockdown, Derivative Assay, In Vitro, Staining, Morphology, Cell Culture, Picogreen Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Analysis, Immunofluorescence, Control

ZDHHC5 overexpression partially counteracts the effects of NOD2 knockdown in human neutrophil-like cells The ability of ZDHHC5 overexpression to counteract the effects of NOD2 knockdown was evaluated in CSE-stimulated, differentiated HL-60-derived neutrophil-like cells. ( A ) Schematic of the in vitro experimental design and treatment groups. ( B ) Representative DAPI-stained images of extracellular DNA structures and quantification of NET formation based on DAPI-stained nuclear morphology. Scale bars: 20 μm. ( C ) Extracellular DNA levels in cell culture supernatants determined using a PicoGreen dsDNA assay. ( D ) MPO levels in cell culture supernatants determined by ELISA. ( E ) NE levels in cell culture supernatants determined by ELISA. ( F ) Representative flow cytometry histograms and quantitative analysis of intracellular ROS levels measured using DCFH-DA staining. ( G ) Representative immunofluorescence images showing LAMP1-positive lysosomes (green) and TOM20-positive mitochondria (red), with nuclei counterstained with DAPI (blue). The colocalization coefficients for LAMP1 and TOM20 were determined by ImageJ and shown on the right. Scale bars: 20 μm. Data are presented as mean ± SD. Data were analyzed using two-way ANOVA followed Tukey’s test. ** p < 0.01 versus the CSE + sh-NC + Vector group; ## p < 0.01 versus the CSE + sh-NOD2 + ZDHHC5 group

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: ZDHHC5 overexpression partially counteracts the effects of NOD2 knockdown in human neutrophil-like cells The ability of ZDHHC5 overexpression to counteract the effects of NOD2 knockdown was evaluated in CSE-stimulated, differentiated HL-60-derived neutrophil-like cells. ( A ) Schematic of the in vitro experimental design and treatment groups. ( B ) Representative DAPI-stained images of extracellular DNA structures and quantification of NET formation based on DAPI-stained nuclear morphology. Scale bars: 20 μm. ( C ) Extracellular DNA levels in cell culture supernatants determined using a PicoGreen dsDNA assay. ( D ) MPO levels in cell culture supernatants determined by ELISA. ( E ) NE levels in cell culture supernatants determined by ELISA. ( F ) Representative flow cytometry histograms and quantitative analysis of intracellular ROS levels measured using DCFH-DA staining. ( G ) Representative immunofluorescence images showing LAMP1-positive lysosomes (green) and TOM20-positive mitochondria (red), with nuclei counterstained with DAPI (blue). The colocalization coefficients for LAMP1 and TOM20 were determined by ImageJ and shown on the right. Scale bars: 20 μm. Data are presented as mean ± SD. Data were analyzed using two-way ANOVA followed Tukey’s test. ** p < 0.01 versus the CSE + sh-NC + Vector group; ## p < 0.01 versus the CSE + sh-NOD2 + ZDHHC5 group

Techniques: Over Expression, Knockdown, Derivative Assay, In Vitro, Staining, Morphology, Cell Culture, Picogreen Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Analysis, Immunofluorescence, Plasmid Preparation

ZDHHC5 overexpression partially counteracts the protective effects of NOD2 knockdown in a mouse model of COPD The ability of ZDHHC5 overexpression to counteract the effects of NOD2 knockdown was evaluated in a cigarette smoke (CS)-induced mouse model of COPD. ( A ) Schematic of the in vivo experimental design. ( B ) qRT-PCR analysis of NOD2 and ZDHHC5 mRNA expression in lung tissue. ( C ) Representative images of H&E staining of lung sections. Scale bar: 100 μm. ( D ) Representative images of Masson’s trichrome staining for collagen deposition. Scale bar: 100 μm. ( E ) Quantification of inflammatory cells and TNF-α, IL-6, and CXCL1 levels in BALF. ( F ) Representative images of IHC staining for E-cadherin and N-cadherin. Scale bar: 50 μm. ( G ) Quantitative analysis of IHC staining. ( H ) Immunofluorescence images showing spatial overlap between Ly6G (green) and LC3B (red). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( I ) Immunofluorescence images showing NETs stained for CitH3 (green) and MPO (red). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( J ) Quantification of CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. Data are presented as mean ± SD. Quantitative data were analyzed using two-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01 vs. COPD + sh-NC + Vector group; # p < 0.05, ## p < 0.01 vs. COPD + sh-NOD2 + ZDHHC5 group

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Figure Lengend Snippet: ZDHHC5 overexpression partially counteracts the protective effects of NOD2 knockdown in a mouse model of COPD The ability of ZDHHC5 overexpression to counteract the effects of NOD2 knockdown was evaluated in a cigarette smoke (CS)-induced mouse model of COPD. ( A ) Schematic of the in vivo experimental design. ( B ) qRT-PCR analysis of NOD2 and ZDHHC5 mRNA expression in lung tissue. ( C ) Representative images of H&E staining of lung sections. Scale bar: 100 μm. ( D ) Representative images of Masson’s trichrome staining for collagen deposition. Scale bar: 100 μm. ( E ) Quantification of inflammatory cells and TNF-α, IL-6, and CXCL1 levels in BALF. ( F ) Representative images of IHC staining for E-cadherin and N-cadherin. Scale bar: 50 μm. ( G ) Quantitative analysis of IHC staining. ( H ) Immunofluorescence images showing spatial overlap between Ly6G (green) and LC3B (red). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( I ) Immunofluorescence images showing NETs stained for CitH3 (green) and MPO (red). Scale bars: 20 μm in the upper panels and 10 μm in the enlarged lower panels. ( J ) Quantification of CitH3-DNA and MPO-DNA complexes in serum and lung tissue homogenates by ELISA. Data are presented as mean ± SD. Quantitative data were analyzed using two-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01 vs. COPD + sh-NC + Vector group; # p < 0.05, ## p < 0.01 vs. COPD + sh-NOD2 + ZDHHC5 group

Techniques: Over Expression, Knockdown, In Vivo, Quantitative RT-PCR, Analysis, Expressing, Tissue, Staining, Immunohistochemistry, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Plasmid Preparation

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Article Snippet: To visualize NETs, mouse lung sections were incubated with antibodies for CitH3 (Cat. # ab232938, Abcam) and myeloperoxidase (MPO) (Cat. #22225-1-AP, Proteintech).

Techniques: Tissue, Immunofluorescence, Marker, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Analysis, Control, Plasmid Preparation

Journal: Biomarker Research

Article Title: ZDHHC5-mediated S-palmitoylation of NOD2 drives neutrophil extracellular trap formation via mitophagy to exacerbate inflammation in COPD

doi: 10.1186/s40364-026-01001-2

Article Snippet: To visualize NETs, mouse lung sections were incubated with antibodies for CitH3 (Cat. # ab232938, Abcam) and myeloperoxidase (MPO) (Cat. #22225-1-AP, Proteintech).

Techniques: Knockdown, Derivative Assay, In Vitro, Staining, Morphology, Cell Culture, Picogreen Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Analysis, Immunofluorescence, Control

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