nox2  (Huabio Inc)

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Suppresses NOX2-Derived ROS to Regulate Macrophage Function and Lesion Progression in End ometriosis (A-B) qPCR analysis of NOX2 mRNA in peritoneal macrophages (PMs) from human (A) and mouse (B) NC and EM groups (n = 3/group). (C-D) Western blot and quantification of NOX2 protein in PBMC-derived macrophages treated with LPS or LPS + 4-OI (n = 5/group). (E) NOX2 enzyme activity in BMDMs (n = 4/group). (F, G) Flow cytometry and quantification of ROS production in BMDMs after LPS or LPS + 4-OI (n = 3/group). (H, I) Flow cytometry and quantification of iNOS + BMDMs after LPS, LPS + 4-OI, or LPS + 4-OI + DPI treatment (n = 4/group). (J) Intracellular Ca 2+ dynamics in BMDMs measured by Fluo-4 after LPS, 4-OI, or DPI treatment. (K) Schematic of 4-OI and DPI intervention in the mouse endometriosis model. (L, M) Representative images (L) and gross morphology (M) of endometriotic lesions after PBS, 4-OI, or DPI treatment. (N) Quantification of lesion weight (n = 6/group). (O, P) Flow cytometry and quantification of iNOS + peritoneal macrophages (percentage and MFI) in peritoneal lavage (n = 6/group). (Q) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages after treatments (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001, ns: not significant.) E2, estradiol benzoate; EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; DPI, NOX2 inhibitor.

Article Snippet: After blocking in 5% milk in TBST, membranes were incubated overnight at 4 °C with primary antibodies: Beta Actin Monoclonal antibody(1:20000, 66009-1-Ig; Proteintech), Beta Tubulin Recombinant antibody (1:5000, 80713-1-RR; Proteintech), Anti-IRG1 antibody (1:1000; ab222411; abcam; Cambridge, UK), Nrf2 monoclonal antibody (1:2000; A21176; abclonal; Wuhan, China), iNOS Polyclonal antibody (1:500; 22226-1-AP; Proteintech), NOX2 Polyclonal antibody (1:3000; 19013-1-AP; Proteintech), p-p38 MAPK Polyclonal antibody (1:2000; 28796-1-AP; Proteintech), and p38 MAPK Polyclonal antibody (1:4000; 14064-1-AP; Proteintech).

Techniques: Derivative Assay, Western Blot, Activity Assay, Flow Cytometry, Control

Journal: Redox Biology

Article Title: A redox sensor in protein kinase A regulatory subunit Iα regulates vasodilation and protects against hypertension

doi: 10.1016/j.redox.2026.104136

Figure Lengend Snippet: Comparison of ex vivo and in vivo vascular responses to angiotensin II acute treatment in wild-type or ‘redox-dead’ Cys17Ser PKARIα knock-in mice. A. Disulfide-PKARIα protein expression in thoracic aortae rings of WT or PKA KI mice subjected either to vehicle, AngII or H 2 O 2 for 5 min (n = 3). B–C. Constriction of abdominal aortae rings from WT or PKARIα KI mice ( B ), or WT or Nox2-null mice ( C ) in response to a single bolus dose of AngII (n ≥ 7). D-E. Acute constriction of abdominal aortae rings from WT or PKA KI mice in response to a single bolus dose of AngII after a vehicle or Nox2-inhibitor GSK2795039 (25 μM, 30 min pre-incubation) pre-treatment ( D ), or a vehicle or Nox1/4-inhibitor GKT137831 (1 μM, 30 min pre-incubation) pre-treatment ( E ) (n ≥ 5). F. Constriction of mesenteric arteries from WT or PKA KI mice in response to a single bolus dose of AngII (n ≥ 5). G. Constriction of penetrating cerebral arterioles in anesthetized WT or PKA KI mice in response to a single bolus administration of AngII (50 μg/kg, i.v.) (n ≥ 7). ∗P< 0.05, ∗∗P< 0.01 vs. vehicle or respective WT. M, PKARIα monomer; D, PKARIα dimer; H 2 O 2 , hydrogen peroxide, acts a vasodilator; AngII, angiotensin II, acts as vasoconstrictor; WT, wild type; PKA KI, Cys17Ser PKARIα knock-in mice; Nox2-null, Nox2 knockout mice; OGB, Oregon Green 488 BAPTA-1, cell permeable calcium dye; Texas Red, dye labels blood vessels in the brain for visualization.

Article Snippet: Nox2−/y (Nox2-null) mice were obtained from Jackson Laboratories [ , ].

Techniques: Comparison, Ex Vivo, In Vivo, Knock-In, Expressing, Incubation, Knock-Out

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Regulates Lysosomal Function, Calcium Signaling, and p38 MAPK Pathway in Macrophages (A) Lysosomal acidification in ascites-derived macrophages from non-endometriosis (NC) and endometriosis (EM) patients assessed by LysoSensor fluorescence intensity (n = 6/group). (B) Lysosomal acidification in PBMC-derived macrophages after LPS,IL-4 and 4-OI treatment (n = 3/group). (C) Lysosomal pH value of BMDMs after LPS and 4-OI treatment (n = 3/group). (D, E) Effect of chloroquine (CQ) on LysoSensor intensity (D) and proportion of iNOS + BMDMs (E) after LPS and 4-OI treatment (n = 3/group). (F) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in BMDMs under indicated treatments (n = 3/group). (G) Intracellular Ca 2+ dynamics in BMDMs after treatments, measured by Fluo-4. (H) Intracellular Ca 2+ dynamics in ascites-derived macrophages from NC and EM patients. (I) Quantification of intracellular calcium in peritoneal macrophages from NC and EM patients (n = 3/group). (J) Quantification of intracellular calcium in BMDMs (n = 5/group). (K) mRNA expression of lysosomal calcium channel genes ( MCOLN1 , MCOLN2 , TPC1 , TPC2 ) in ascites-derived macrophages from NC and EM patients (n = 3/group). (L) lysosomal calcium channel genes mRNA in PBMC-derived macrophages co-cultured with normal ESCs (Nor-ESC), eutopic ESCs (Eu-ESC), or ectopic ESCs (Ec-ESC) from patients (n = 5/group). (M − N) Fluo-4-based Ca 2+ dynamics in BMDMs treated with LPS, 4-OI, ML-SA1 (M), or CQ (N). (O) Flow cytometry analysis and quantification of iNOS + BMDMs after indicated treatments (n = 3/group). (P) mRNA levels of pro-inflammatory genes in BMDMs under different treatments (n = 3/group). (Q) Western blot and quantification of p-p38 and total p38 in BMDMs with LPS ± 4-OI (n = 3/group). (R–S) Western blot and quantification of iNOS, p-p38, and total p38 in BMDMs treated with LPS, 4-OI, and CQ (R), or ML-SA1 (S) (n = 3/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; CQ, chloroquine; ML-SA1, MCOLN channel agonist; PBMC, peripheral blood mononuclear cell; ESC, endometrial stromal cell.

Article Snippet: After blocking in 5% milk in TBST, membranes were incubated overnight at 4 °C with primary antibodies: Beta Actin Monoclonal antibody(1:20000, 66009-1-Ig; Proteintech), Beta Tubulin Recombinant antibody (1:5000, 80713-1-RR; Proteintech), Anti-IRG1 antibody (1:1000; ab222411; abcam; Cambridge, UK), Nrf2 monoclonal antibody (1:2000; A21176; abclonal; Wuhan, China), iNOS Polyclonal antibody (1:500; 22226-1-AP; Proteintech), NOX2 Polyclonal antibody (1:3000; 19013-1-AP; Proteintech), p-p38 MAPK Polyclonal antibody (1:2000; 28796-1-AP; Proteintech), and p38 MAPK Polyclonal antibody (1:4000; 14064-1-AP; Proteintech).

Techniques: Derivative Assay, Fluorescence, Expressing, Cell Culture, Flow Cytometry, Western Blot, Control

Journal: Current Issues in Molecular Biology

Article Title: Coptis chinensis Franch. Suppresses Invasive Pulmonary Aspergillosis by Augmenting NADPH-Dependent Neutrophil Extracellular Traps via Dual Modulation of Complement Activation and Gut Microbiota

doi: 10.3390/cimb48040424

Figure Lengend Snippet: Bayesian networks and KEGG suggest a CCF-biased reorganization toward CR3-SYK-NOX/NET pathways relative to IPA/AmB. The network analysis highlights recurrent CCF-associated hub genes, including Itgam, Syk, Rac2, and Cybb, together with pathway enrichment linked to oxidative and NET-related responses. ( A ) Hub gene networks identified in the IPA, CCF, and AmB groups by Bayesian network analysis. Genes were ranked by DEGREE value, with the top 10% defined as hub genes and marked in color; nodes with DEGREE >6 were placed at the core of the network. ( B ) KEGG pathway enrichment of hub genes in each group. Pathways were ranked by −log 10 ( p value). Bar colors represent different groups: IPA (dark blue), CCF (dark red), and AmB (light blue). The red box denotes the most significantly enriched KEGG pathway in the CCF group. ( C ) FPKM values of key CCF-regulated genes at +2, +4, and +6 dpi. Statistical analysis was conducted comparing all groups to the CCF group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 indicate levels of statistical significance. ( D ) qPCR validation of key CCF-regulated genes in each group. Statistical analysis was conducted using one-way ANOVA, comparing all groups to the CCF group. * p < 0.05, ** p < 0.01, **** p < 0.0001 indicate levels of statistical significance. Data are presented as mean ± SD from n = 3 mice per group.

Article Snippet: Primary antibodies used in Western Blot included Beta Actin Monoclonal antibody (66009-1-Ig, Proteintech, Wuhan, China), NCF4/p40phox Rabbit pAb (A2096, Abclonal, Wuhan, China), Anti-NOX2/gp91phox Recombinant Rabbit Monoclonal Antibody (ET1611-44, Huabio, Hangzhou, China), RAC2 Rabbit pAb (A1139, Abclonal, China) and Anti-NCF1/p47phox Rabbit pAb (GB11724, Servicebio, Wuhan, China).

Techniques: Biomarker Discovery

Journal: Current Issues in Molecular Biology

Article Title: Coptis chinensis Franch. Suppresses Invasive Pulmonary Aspergillosis by Augmenting NADPH-Dependent Neutrophil Extracellular Traps via Dual Modulation of Complement Activation and Gut Microbiota

doi: 10.3390/cimb48040424

Figure Lengend Snippet: CCF enhances NOX2 components and its benefit is partially attenuated by apocynin, consistent with oxidative pathway involvement. This figure combines protein, survival, and fungal-burden data to show partial pharmacological support for the role of oxidative responses in CCF-mediated protection. ( A ) Protein expression of NADPH oxidase subunits in lung tissue at +4 dpi as detected by Western blot. I, IPA group; C, CCF group; A, AmB group; M, Marker. Data are presented as mean ± SD from n = 5 mice per group. ( B ) Densitometric quantification of protein levels normalized to β-ACTIN. Statistical analysis was performed using one-way ANOVA with multiple comparisons. * p < 0.05. Data are presented as mean ± SD from n = 5 mice per group. ( C ) Survival curves of mice up to +8 dpi. CCF significantly improved survival compared to the IPA group. Co-treatment with the NADPH oxidase inhibitor apocynin partially reversed this effect. * p < 0.05, ** p < 0.01. Survival data were analyzed using the Kaplan–Meier method with n = 10 mice per group. ( D ) Fungal burden in lung tissues at +4 dpi measured by colony-forming unit counts. Statistical analysis was performed using one-way ANOVA with multiple comparisons. * p < 0.05. Data are presented as mean ± SD from n = 5 mice per group.

Article Snippet: Primary antibodies used in Western Blot included Beta Actin Monoclonal antibody (66009-1-Ig, Proteintech, Wuhan, China), NCF4/p40phox Rabbit pAb (A2096, Abclonal, Wuhan, China), Anti-NOX2/gp91phox Recombinant Rabbit Monoclonal Antibody (ET1611-44, Huabio, Hangzhou, China), RAC2 Rabbit pAb (A1139, Abclonal, China) and Anti-NCF1/p47phox Rabbit pAb (GB11724, Servicebio, Wuhan, China).

Techniques: Expressing, Western Blot, Marker

Journal: Current Issues in Molecular Biology

Article Title: Coptis chinensis Franch. Suppresses Invasive Pulmonary Aspergillosis by Augmenting NADPH-Dependent Neutrophil Extracellular Traps via Dual Modulation of Complement Activation and Gut Microbiota

doi: 10.3390/cimb48040424

Figure Lengend Snippet: CCF-treated mice show microbiota remodeling with enrichment of Clostridium sp., and observed Cybb/Syk associations are non-causal. The main features of this figure are the distinct clustering of gut microbial communities across groups, the selective enrichment of Clostridium sp. in the CCF group, and its positive correlation with the redox-related host genes Cybb and Syk . ( A ) Alpha diversity analysis using the Chao1 index and Shannon index reveals significant differences among CN, IPA, CCF, and AmB groups. ( B ) Principal coordinates analysis (PCoA) based on weighted UniFrac distances demonstrates distinct clustering of gut microbial communities across groups. ( C ) Taxonomic composition profiles at the family genus, and species levels. Bar plots display the relative abundance of dominant taxa within each group. ( D ) Histogram of LDA scores (log 10 ) for discriminative features, indicating significantly enriched taxa in each group. The colors represent different groups: CN (blue), IPA (red), CCF (green), and AmB (purple). ( E ) Cladogram representing the phylogenetic distribution of differentially abundant taxa among the groups. Colored nodes represent taxa significantly enriched in the corresponding groups: CN (blue), IPA (red), CCF (green), and AmB (purple). From inner to outer circles represent phylum, class, order, family, genus, and species levels. Taxa connected by the same color belong to the same species. ( F ) Comparison of the abundances of significantly different biomarkers: g__ Parasutterella and s__ Lactobacillus in the CN group, s__ Clostridium_sp in the CCF group, g__ Clostridia in the AmB group, and significantly deleted biomarkers g__ Muribaculaceae and g__ Lachnospiraceae in the CCF group. ( G ) Spearman correlation analysis between Clostridium sp. abundance and the expression levels of NADPH oxidase-related genes. Scatter plots show strong positive correlations with Cybb (R = 0.81, p = 0.0076) and Syk (R = 0.75, p = 0.021).

Article Snippet: Primary antibodies used in Western Blot included Beta Actin Monoclonal antibody (66009-1-Ig, Proteintech, Wuhan, China), NCF4/p40phox Rabbit pAb (A2096, Abclonal, Wuhan, China), Anti-NOX2/gp91phox Recombinant Rabbit Monoclonal Antibody (ET1611-44, Huabio, Hangzhou, China), RAC2 Rabbit pAb (A1139, Abclonal, China) and Anti-NCF1/p47phox Rabbit pAb (GB11724, Servicebio, Wuhan, China).

Techniques: Comparison, Expressing