Structured Review

Proteintech anti gp91 phox
a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress <t>GP91-PHOX</t> (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).
Anti Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti gp91 phox/product/Proteintech
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti gp91 phox - by Bioz Stars, 2024-05
86/100 stars

Images

1) Product Images from "Mitochondrial complex I activity in microglia sustains neuroinflammation"

Article Title: Mitochondrial complex I activity in microglia sustains neuroinflammation

Journal: Nature

doi: 10.1038/s41586-024-07167-9

a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress GP91-PHOX (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).
Figure Legend Snippet: a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress GP91-PHOX (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).

Techniques Used: Transformation Assay, Expressing, Marker

a , Seahorse metabolic flux analysis of primary microglia derived from wild-type (WT) and Nd6 mice under basal conditions and after stimulation with LPS and IFNγ. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. Differences in basal respiration, mitochondrial (mt) ATP production and maximal respiration are reported. b , Quantification of mtROS production in LPS + IFNγ-stimulated (pro-inflammatory) primary WT and Nd6 microglia after RET induction (RET + ). From left to right, n = 18, 18, 17, 18, 18 and 18 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. c , Quantification of neuronal neurite length after co-culture with RET + pro-inflammatory primary WT and Nd6 microglia. From left to right, n = 11, 5, 6, 12, 12, 12 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. d , EAE scores of WT and Nd6 mice up to 30 days after immunization. n = 17 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. e , f , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 13,614 cells (7,501 (WT) and 6,113 (Nd6)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( e ) and EAE ( f ) mice (30 days after immunization). g , Selected hMG-like and DAM genes in cluster 0 and 1 DAMs. h , The mitochondrial membrane potential (Δ ψ m ) in ex vivo FACS-isolated CD45 + CD11b + cells. From left to right, n = 4, 3, 4 and 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Fisher’s LSD test. i , j Representative images and quantifications of perilesional microglial branching ( i ; n = 12 replicates per group) and IBA1 + SPP1 + GP91-PHOX + cells in WT and Nd6 EAE mice ( j ; n = 4 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. For i and j , scale bars, 30 μm. For d , i and j , data are mean ± s.e.m. The violin plots in a – c , and h show the median and quartiles. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Figure Legend Snippet: a , Seahorse metabolic flux analysis of primary microglia derived from wild-type (WT) and Nd6 mice under basal conditions and after stimulation with LPS and IFNγ. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. Differences in basal respiration, mitochondrial (mt) ATP production and maximal respiration are reported. b , Quantification of mtROS production in LPS + IFNγ-stimulated (pro-inflammatory) primary WT and Nd6 microglia after RET induction (RET + ). From left to right, n = 18, 18, 17, 18, 18 and 18 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. c , Quantification of neuronal neurite length after co-culture with RET + pro-inflammatory primary WT and Nd6 microglia. From left to right, n = 11, 5, 6, 12, 12, 12 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. d , EAE scores of WT and Nd6 mice up to 30 days after immunization. n = 17 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. e , f , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 13,614 cells (7,501 (WT) and 6,113 (Nd6)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( e ) and EAE ( f ) mice (30 days after immunization). g , Selected hMG-like and DAM genes in cluster 0 and 1 DAMs. h , The mitochondrial membrane potential (Δ ψ m ) in ex vivo FACS-isolated CD45 + CD11b + cells. From left to right, n = 4, 3, 4 and 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Fisher’s LSD test. i , j Representative images and quantifications of perilesional microglial branching ( i ; n = 12 replicates per group) and IBA1 + SPP1 + GP91-PHOX + cells in WT and Nd6 EAE mice ( j ; n = 4 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. For i and j , scale bars, 30 μm. For d , i and j , data are mean ± s.e.m. The violin plots in a – c , and h show the median and quartiles. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Techniques Used: Derivative Assay, Co-Culture Assay, Membrane, Ex Vivo, Isolation, Two Tailed Test

a , EAE scores of Ndufs4- WT and Ndufs4 -KO mice. n = 10 (WT) and 12 (KO) mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. b , c , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 10,666 cells (4,180 ( Ndufs4 WT) and 6,486 ( Ndufs4 KO)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( b ) and EAE ( c ) mice (50 days after immunization). d , Selected hMG-like and DAM genes in cluster 2 and 6 DAMs. e , Suspension mass cytometry (CyTOF) analysis of immune cell types at 50 days after immunization obtained from 51,177 cells (23,467 ( Ndufs4 WT); 27,710 ( Ndufs4 KO)). AA, alternatively activated; pro-inflam., pro-inflammatory. f , g , Quantification of CX3CR1 + SPP1 + ( f ; n = 5 replicates per group) and CASPASE3 + IBA1 + ( g ; n = 4 replicates per group) cells in EAE. Statistical analysis was performed using two-tailed unpaired t -tests. h – j , In vivo quantification of perilesional microglial branching ( h ; n = 12 (WT) and 11 (KO) replicates per group; two-tailed unpaired t -test), GP91-PHOX expression in the EAE spinal cords ( i ; n = 5 (WT) and 6 (KO) replicates per group; two-tailed Mann–Whitney U -test) and IBA1 + SPP1 + GP91-PHOX + cells ( j ; n = 4 replicates per group; two-tailed unpaired t -test). Scale bars, 7 μm ( h ) and 400 μm ( i ). k , l , Representative images and quantification of axonal loss ( k ; n = 5 (WT) and 6 (KO) replicates per group) and axonal degeneration ( l ; n = 5 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. APP, amyloid precursor protein; NHP, neurofilament heavy polypeptide. Insets: merged images. Scale bars, 400 μm ( k ) and 50 μm ( l ). m , EAE scores of mice treated with metformin, DMM, DMM + metformin versus saline controls. n = 13 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction; # P < 0.05 comparing DMM + metformin versus saline. n , CyTOF analysis of immune cell types at 30 days after immunization obtained from 159,110 cells (32,793 (metformin), 40,864 (DMM), 44,143 (DMM + metformin) and 41,310 (saline)). o , Quantification of CX3CR1 + SPP1 + NDUFS4 + cells, oxidative stress, axonal loss and axonal degeneration in EAE mice. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. For a , f – m and o , data are mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, *** P < 0.0001.
Figure Legend Snippet: a , EAE scores of Ndufs4- WT and Ndufs4 -KO mice. n = 10 (WT) and 12 (KO) mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. b , c , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 10,666 cells (4,180 ( Ndufs4 WT) and 6,486 ( Ndufs4 KO)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( b ) and EAE ( c ) mice (50 days after immunization). d , Selected hMG-like and DAM genes in cluster 2 and 6 DAMs. e , Suspension mass cytometry (CyTOF) analysis of immune cell types at 50 days after immunization obtained from 51,177 cells (23,467 ( Ndufs4 WT); 27,710 ( Ndufs4 KO)). AA, alternatively activated; pro-inflam., pro-inflammatory. f , g , Quantification of CX3CR1 + SPP1 + ( f ; n = 5 replicates per group) and CASPASE3 + IBA1 + ( g ; n = 4 replicates per group) cells in EAE. Statistical analysis was performed using two-tailed unpaired t -tests. h – j , In vivo quantification of perilesional microglial branching ( h ; n = 12 (WT) and 11 (KO) replicates per group; two-tailed unpaired t -test), GP91-PHOX expression in the EAE spinal cords ( i ; n = 5 (WT) and 6 (KO) replicates per group; two-tailed Mann–Whitney U -test) and IBA1 + SPP1 + GP91-PHOX + cells ( j ; n = 4 replicates per group; two-tailed unpaired t -test). Scale bars, 7 μm ( h ) and 400 μm ( i ). k , l , Representative images and quantification of axonal loss ( k ; n = 5 (WT) and 6 (KO) replicates per group) and axonal degeneration ( l ; n = 5 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. APP, amyloid precursor protein; NHP, neurofilament heavy polypeptide. Insets: merged images. Scale bars, 400 μm ( k ) and 50 μm ( l ). m , EAE scores of mice treated with metformin, DMM, DMM + metformin versus saline controls. n = 13 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction; # P < 0.05 comparing DMM + metformin versus saline. n , CyTOF analysis of immune cell types at 30 days after immunization obtained from 159,110 cells (32,793 (metformin), 40,864 (DMM), 44,143 (DMM + metformin) and 41,310 (saline)). o , Quantification of CX3CR1 + SPP1 + NDUFS4 + cells, oxidative stress, axonal loss and axonal degeneration in EAE mice. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. For a , f – m and o , data are mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, *** P < 0.0001.

Techniques Used: Suspension, Mass Cytometry, Two Tailed Test, In Vivo, Expressing, MANN-WHITNEY, Saline

a , Cytotoxicity assay and mtROS production of mouse BV2 microglia treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 7, 3, 3, 3, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (first column, top-bottom), n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 37, 11, 14, 11, 11, 10, 11, 6, 6, 6, 3, 11, 11 replicates per group (first column, top-bottom), n = 46, 11, 14, 11, 11, 18, 17, 5, 6, 6, 3, 18, 18 replicates per group (second column, top-bottom), n = 38, 20, 16, 20, 20, 18, 18, 8, 8, 8, 4, 18, 18 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. b , Cytotoxicity assay and mtROS production of human induced microglia (hiMG) treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (first column, top-bottom), n = 7, 4, 3, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 27, 10, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (first column, top-bottom), n = 32, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (second column, top-bottom), n = 31, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 5 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. c , EAE scores of mice treated with 4-octyl itaconate (4-OI) vs saline controls (n = 6 mice per group; mean ± SEM; two-way ANOVA; Bonferroni). d , e , In vivo testing of dimethyl malonate (DMM) and disodium malonate (DSM) in EAE. ( d ) EAE mice receiving daily intraperitoneal (IP) injections of DMM (160 mg/kg), DSM (160 mg/kg), or saline at 7 days from disease onset. Malonate levels in the peripheral blood (at 30 min from injection, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). ***P < 0.001. ( e ) EAE mice receiving DMM (1.5%), DSM (1.5%), or saline dissolved in their drinking water (at 7 days from disease onset). Malonate levels in the peripheral blood during treatment (daily, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). *P < 0.05, **P < 0.01. f , UMAP plot from the CyTOF data at 30 dpi (see also Fig. ) coloured by cell type found in EAE mice treated with CII and CI inhibitors. Numbers of clusters are shown in superimposition. A.A.: alternatively activated. g , Fraction of cell clusters in EAE mice treated with metformin, DMM, DMM+metformin, vs saline-treated controls at 30 dpi. h , i , Expression of ( h ) CII and ( i ) CI in the CyTOF clusters identified as hMG-like cells and DAM (median and quartiles from 159,110 cells; *P < 0.01, **P < 0.0001; unpaired t-test, two-tailed). j , Representative confocal images and quantification of the expression of the marker of oxidative stress GP91-PHOX in microglia (IBA1 + ), astrocytes (GFAP + ), oligodendrocytes (OLIG2 + ), and neurons (NEUN + ) in the spinal cord of treated EAE mice (n = 4 replicates per group; mean ± SEM; *P < 0.05; one-way ANOVA; Tukey). Abbreviations: rotenone (rot.), metformin (met.), 4-octyl itaconate (4-OI), dimethyl malonate (DMM), disodium malonate (DSM).
Figure Legend Snippet: a , Cytotoxicity assay and mtROS production of mouse BV2 microglia treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 7, 3, 3, 3, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (first column, top-bottom), n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 37, 11, 14, 11, 11, 10, 11, 6, 6, 6, 3, 11, 11 replicates per group (first column, top-bottom), n = 46, 11, 14, 11, 11, 18, 17, 5, 6, 6, 3, 18, 18 replicates per group (second column, top-bottom), n = 38, 20, 16, 20, 20, 18, 18, 8, 8, 8, 4, 18, 18 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. b , Cytotoxicity assay and mtROS production of human induced microglia (hiMG) treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (first column, top-bottom), n = 7, 4, 3, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 27, 10, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (first column, top-bottom), n = 32, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (second column, top-bottom), n = 31, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 5 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. c , EAE scores of mice treated with 4-octyl itaconate (4-OI) vs saline controls (n = 6 mice per group; mean ± SEM; two-way ANOVA; Bonferroni). d , e , In vivo testing of dimethyl malonate (DMM) and disodium malonate (DSM) in EAE. ( d ) EAE mice receiving daily intraperitoneal (IP) injections of DMM (160 mg/kg), DSM (160 mg/kg), or saline at 7 days from disease onset. Malonate levels in the peripheral blood (at 30 min from injection, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). ***P < 0.001. ( e ) EAE mice receiving DMM (1.5%), DSM (1.5%), or saline dissolved in their drinking water (at 7 days from disease onset). Malonate levels in the peripheral blood during treatment (daily, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). *P < 0.05, **P < 0.01. f , UMAP plot from the CyTOF data at 30 dpi (see also Fig. ) coloured by cell type found in EAE mice treated with CII and CI inhibitors. Numbers of clusters are shown in superimposition. A.A.: alternatively activated. g , Fraction of cell clusters in EAE mice treated with metformin, DMM, DMM+metformin, vs saline-treated controls at 30 dpi. h , i , Expression of ( h ) CII and ( i ) CI in the CyTOF clusters identified as hMG-like cells and DAM (median and quartiles from 159,110 cells; *P < 0.01, **P < 0.0001; unpaired t-test, two-tailed). j , Representative confocal images and quantification of the expression of the marker of oxidative stress GP91-PHOX in microglia (IBA1 + ), astrocytes (GFAP + ), oligodendrocytes (OLIG2 + ), and neurons (NEUN + ) in the spinal cord of treated EAE mice (n = 4 replicates per group; mean ± SEM; *P < 0.05; one-way ANOVA; Tukey). Abbreviations: rotenone (rot.), metformin (met.), 4-octyl itaconate (4-OI), dimethyl malonate (DMM), disodium malonate (DSM).

Techniques Used: Cytotoxicity Assay, In Vitro, Two Tailed Test, Saline, In Vivo, Injection, Expressing, Marker


Structured Review

Proteintech gp91 phox antibodies
Gp91 Phox Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gp91 phox antibodies/product/Proteintech
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gp91 phox antibodies - by Bioz Stars, 2024-05
86/100 stars

Images


Structured Review

Proteintech anti gp91 phox
Anti Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti gp91 phox/product/Proteintech
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti gp91 phox - by Bioz Stars, 2024-05
94/100 stars

Images


Structured Review

Proteintech gp91 phox
Treatment of LXA4 inhibits oxidative stress in corpus cavernous of DMED rats. ( a ) Representative Western blot results for the p22 <t>phox</t> , p47 phox , <t>gp91</t> phox , p40 phox , and p67 phox in rats of all the three groups. ( b ) Expressions of p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox with β-actin as the loading control in all the three groups were presented through bar graphs. ( c ) MDA levels determined by the ELISA method in all the three groups. ( d ) SOD activities determined by the ELISA method in all the three groups. Data are expressed as mean ± s.e.m. ( n control = 10, n DMED = 11, and n DMED + LXA4 = 11). * P < 0.05 and *** P < 0.001 when comparing two groups. DMED: diabetes mellitus-induced erectile dysfunction; LXA4: Lipoxin A4; ELISA: enzyme-linked immunosorbent assay; SOD: superoxide dismutase; MDA: malondialdehyde; s.e.m.: standard error of the mean.
Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gp91 phox/product/Proteintech
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gp91 phox - by Bioz Stars, 2024-05
94/100 stars

Images

1) Product Images from "Lipoxin A4 improves erectile dysfunction in rats with type I diabetes by inhibiting oxidative stress and corporal fibrosis"

Article Title: Lipoxin A4 improves erectile dysfunction in rats with type I diabetes by inhibiting oxidative stress and corporal fibrosis

Journal: Asian Journal of Andrology

doi: 10.4103/aja.aja_49_17

Treatment of LXA4 inhibits oxidative stress in corpus cavernous of DMED rats. ( a ) Representative Western blot results for the p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox in rats of all the three groups. ( b ) Expressions of p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox with β-actin as the loading control in all the three groups were presented through bar graphs. ( c ) MDA levels determined by the ELISA method in all the three groups. ( d ) SOD activities determined by the ELISA method in all the three groups. Data are expressed as mean ± s.e.m. ( n control = 10, n DMED = 11, and n DMED + LXA4 = 11). * P < 0.05 and *** P < 0.001 when comparing two groups. DMED: diabetes mellitus-induced erectile dysfunction; LXA4: Lipoxin A4; ELISA: enzyme-linked immunosorbent assay; SOD: superoxide dismutase; MDA: malondialdehyde; s.e.m.: standard error of the mean.
Figure Legend Snippet: Treatment of LXA4 inhibits oxidative stress in corpus cavernous of DMED rats. ( a ) Representative Western blot results for the p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox in rats of all the three groups. ( b ) Expressions of p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox with β-actin as the loading control in all the three groups were presented through bar graphs. ( c ) MDA levels determined by the ELISA method in all the three groups. ( d ) SOD activities determined by the ELISA method in all the three groups. Data are expressed as mean ± s.e.m. ( n control = 10, n DMED = 11, and n DMED + LXA4 = 11). * P < 0.05 and *** P < 0.001 when comparing two groups. DMED: diabetes mellitus-induced erectile dysfunction; LXA4: Lipoxin A4; ELISA: enzyme-linked immunosorbent assay; SOD: superoxide dismutase; MDA: malondialdehyde; s.e.m.: standard error of the mean.

Techniques Used: Western Blot, Enzyme-linked Immunosorbent Assay


Structured Review

Proteintech anti gp91 phox
Anti Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti gp91 phox/product/Proteintech
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti gp91 phox - by Bioz Stars, 2024-05
94/100 stars

Images


Structured Review

Proteintech gp91 phox
GDF11 inhibited oxidative damage in diabetic cardiomyopathy (DCM) by upregulating SIRT1 expression. (A) Representative images of dihydroethidium (DHE) staining (scale bar = 50 μm). (B) DHE fluorescence intensities. (C) Malondialdehyde (MDA) content. (D) Superoxide dismutase (SOD) activity. (E) Glutathione peroxidase (GSH-Px) activity. (F) Representative images of SIRT1 immunofluorescence in heart tissue. (G) The deacetylase activity of SIRT1. (H) Representative blots of SIRT1, Nrf2, <t>gp91</t> <t>phox</t> , SOD2, and HO1. (I) Quantitative expression of SIRT1. (J) Quantitative expression of Nrf2. (K) Quantitative expression of gp91 phox . (L) Quantitative expression of SOD2. (M) Quantitative expression of HO1. Data are presented as the mean ± SEM, n = 5 or 6 per group. *,** P < 0.05, 0.01 versus the Con group, †† P < 0.01 versus the DM group.
Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gp91 phox/product/Proteintech
Average 92 stars, based on 1 article reviews
Price from $9.99 to $1999.99
gp91 phox - by Bioz Stars, 2024-05
92/100 stars

Images

1) Product Images from "GDF11 Alleviates Pathological Myocardial Remodeling in Diabetic Cardiomyopathy Through SIRT1-Dependent Regulation of Oxidative Stress and Apoptosis"

Article Title: GDF11 Alleviates Pathological Myocardial Remodeling in Diabetic Cardiomyopathy Through SIRT1-Dependent Regulation of Oxidative Stress and Apoptosis

Journal: Frontiers in Cell and Developmental Biology

doi: 10.3389/fcell.2021.686848

GDF11 inhibited oxidative damage in diabetic cardiomyopathy (DCM) by upregulating SIRT1 expression. (A) Representative images of dihydroethidium (DHE) staining (scale bar = 50 μm). (B) DHE fluorescence intensities. (C) Malondialdehyde (MDA) content. (D) Superoxide dismutase (SOD) activity. (E) Glutathione peroxidase (GSH-Px) activity. (F) Representative images of SIRT1 immunofluorescence in heart tissue. (G) The deacetylase activity of SIRT1. (H) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (I) Quantitative expression of SIRT1. (J) Quantitative expression of Nrf2. (K) Quantitative expression of gp91 phox . (L) Quantitative expression of SOD2. (M) Quantitative expression of HO1. Data are presented as the mean ± SEM, n = 5 or 6 per group. *,** P < 0.05, 0.01 versus the Con group, †† P < 0.01 versus the DM group.
Figure Legend Snippet: GDF11 inhibited oxidative damage in diabetic cardiomyopathy (DCM) by upregulating SIRT1 expression. (A) Representative images of dihydroethidium (DHE) staining (scale bar = 50 μm). (B) DHE fluorescence intensities. (C) Malondialdehyde (MDA) content. (D) Superoxide dismutase (SOD) activity. (E) Glutathione peroxidase (GSH-Px) activity. (F) Representative images of SIRT1 immunofluorescence in heart tissue. (G) The deacetylase activity of SIRT1. (H) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (I) Quantitative expression of SIRT1. (J) Quantitative expression of Nrf2. (K) Quantitative expression of gp91 phox . (L) Quantitative expression of SOD2. (M) Quantitative expression of HO1. Data are presented as the mean ± SEM, n = 5 or 6 per group. *,** P < 0.05, 0.01 versus the Con group, †† P < 0.01 versus the DM group.

Techniques Used: Expressing, Staining, Fluorescence, Activity Assay, Immunofluorescence, Histone Deacetylase Assay

SIRT1 inhibition abolished the antioxidative and antiapoptotic functions of GDF11 in diabetic cardiomyopathy (DCM). (A) Representative images of dihydroethidium (DHE) staining (scale bar = 50 μm). (B) DHE fluorescence intensities. (C) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (D) Quantitative expression of SIRT1. (E) Quantitative expression of Nrf2. (F) Quantitative expression of gp91 phox . (G) Quantitative expression of SOD2. (H) Quantitative expression of HO1. (I) Representative images of TUNEL staining (scale bar = 50 μm). (J) Apoptotic ratio. (K) Representative blots of p-p65. p65, Bax, Bcl-2, and Cleaved Caspase-3. (L) Quantitative analysis of the ratio of p-p65 to p65. (M) Quantitative expression of Bax. (N) Quantitative expression of Bcl-2. (O) Quantitative expression of Cleaved Caspase-3. Data are presented as the mean ± SEM, n = 5 or 6 per group. ** P < 0.01 versus the DM group, †† P < 0.01 versus the DM + AAV-GDF11 group.
Figure Legend Snippet: SIRT1 inhibition abolished the antioxidative and antiapoptotic functions of GDF11 in diabetic cardiomyopathy (DCM). (A) Representative images of dihydroethidium (DHE) staining (scale bar = 50 μm). (B) DHE fluorescence intensities. (C) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (D) Quantitative expression of SIRT1. (E) Quantitative expression of Nrf2. (F) Quantitative expression of gp91 phox . (G) Quantitative expression of SOD2. (H) Quantitative expression of HO1. (I) Representative images of TUNEL staining (scale bar = 50 μm). (J) Apoptotic ratio. (K) Representative blots of p-p65. p65, Bax, Bcl-2, and Cleaved Caspase-3. (L) Quantitative analysis of the ratio of p-p65 to p65. (M) Quantitative expression of Bax. (N) Quantitative expression of Bcl-2. (O) Quantitative expression of Cleaved Caspase-3. Data are presented as the mean ± SEM, n = 5 or 6 per group. ** P < 0.01 versus the DM group, †† P < 0.01 versus the DM + AAV-GDF11 group.

Techniques Used: Inhibition, Staining, Fluorescence, Expressing, TUNEL Assay

GDF11 overexpression attenuated high glucose (HG) and palmitate (Pal) induced reactive oxygen species (ROS) overproduction in H9c2 cells. (A) Representative images of 2’,7’-dichlorofluorescein diacetate (DCFH-DA) staining (scale bar = 200 μm). (B) DCFH-DA fluorescence intensities. (C) The deacetylase activity of SIRT1. (D) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (E) Quantitative expression of SIRT1. (F) Quantitative expression of Nrf2. (G) Quantitative expression of gp91 phox . (H) Quantitative expression of SOD2. (I) Quantitative expression of HO1. Data are presented as the mean ± SEM, n = 5 or 6 per group. #/## P < 0.05/0.01 versus the NG group, $$ P < 0.01 versus the HG + Pal group.
Figure Legend Snippet: GDF11 overexpression attenuated high glucose (HG) and palmitate (Pal) induced reactive oxygen species (ROS) overproduction in H9c2 cells. (A) Representative images of 2’,7’-dichlorofluorescein diacetate (DCFH-DA) staining (scale bar = 200 μm). (B) DCFH-DA fluorescence intensities. (C) The deacetylase activity of SIRT1. (D) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (E) Quantitative expression of SIRT1. (F) Quantitative expression of Nrf2. (G) Quantitative expression of gp91 phox . (H) Quantitative expression of SOD2. (I) Quantitative expression of HO1. Data are presented as the mean ± SEM, n = 5 or 6 per group. #/## P < 0.05/0.01 versus the NG group, $$ P < 0.01 versus the HG + Pal group.

Techniques Used: Over Expression, Staining, Fluorescence, Histone Deacetylase Assay, Activity Assay, Expressing

SIRT1 siRNA diminished GDF11-induced inhibition of oxidative stress and apoptosis in H9c2 cells. (A) Representative images of 2’,7’-dichlorofluorescein diacetate (DCFH-DA) staining (scale bar = 200 μm). (B) DCFH-DA fluorescence intensities. (C) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (D) Quantitative expression of SIRT1. (E) Quantitative expression of Nrf2. (F) Quantitative expression of gp91 phox . (G) Quantitative expression of SOD2. (H) Quantitative expression of HO1. (I) Representative images of TUNEL staining (scale bar = 200 μm). (J) Apoptotic ratio. (K) Representative blots of p-p65, p65, Bax, Bcl-2, and Cleaved Caspase-3. (L) Quantitative analysis of the ratio of p-p65 to p65. (M) Quantitative expression of Bax. (N) Quantitative expression of Bcl-2. (O) Quantitative expression of Cleaved Caspase-3. Data are presented as the mean ± SEM, n = 5 or 6 per group. ## P < 0.01 versus the HG + Pal group, $$ P < 0.01 versus the HG + Pal + Ad-GDF11 group.
Figure Legend Snippet: SIRT1 siRNA diminished GDF11-induced inhibition of oxidative stress and apoptosis in H9c2 cells. (A) Representative images of 2’,7’-dichlorofluorescein diacetate (DCFH-DA) staining (scale bar = 200 μm). (B) DCFH-DA fluorescence intensities. (C) Representative blots of SIRT1, Nrf2, gp91 phox , SOD2, and HO1. (D) Quantitative expression of SIRT1. (E) Quantitative expression of Nrf2. (F) Quantitative expression of gp91 phox . (G) Quantitative expression of SOD2. (H) Quantitative expression of HO1. (I) Representative images of TUNEL staining (scale bar = 200 μm). (J) Apoptotic ratio. (K) Representative blots of p-p65, p65, Bax, Bcl-2, and Cleaved Caspase-3. (L) Quantitative analysis of the ratio of p-p65 to p65. (M) Quantitative expression of Bax. (N) Quantitative expression of Bcl-2. (O) Quantitative expression of Cleaved Caspase-3. Data are presented as the mean ± SEM, n = 5 or 6 per group. ## P < 0.01 versus the HG + Pal group, $$ P < 0.01 versus the HG + Pal + Ad-GDF11 group.

Techniques Used: Inhibition, Staining, Fluorescence, Expressing, TUNEL Assay


Structured Review

Proteintech gp91 phox
Analysis of the expression levels of <t>gp91-phox</t> and SOD1. ( A and B ) The expression levels of gp91-phox and SOD1 analyzed by Western Blot in LSCs and Non-LSCs treated with different agents. ( C – G ) Semi-quantification of the expression levels of gp91-phox, SOD1, CD38, DEPTOR, and IFITM3 detected by RT-qPCR in LSCs and Non-LSCs treated with different agents. * p <0.05, ** p <0.01 and *** p < 0.001 were calculated by t test, referring to the statistically significant difference as compared to respective group.
Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Iron Oxide Nanoparticles Combined with Cytosine Arabinoside Show Anti-Leukemia Stem Cell Effects on Acute Myeloid Leukemia by Regulating Reactive Oxygen Species"

Article Title: Iron Oxide Nanoparticles Combined with Cytosine Arabinoside Show Anti-Leukemia Stem Cell Effects on Acute Myeloid Leukemia by Regulating Reactive Oxygen Species

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S278885

Analysis of the expression levels of gp91-phox and SOD1. ( A and B ) The expression levels of gp91-phox and SOD1 analyzed by Western Blot in LSCs and Non-LSCs treated with different agents. ( C – G ) Semi-quantification of the expression levels of gp91-phox, SOD1, CD38, DEPTOR, and IFITM3 detected by RT-qPCR in LSCs and Non-LSCs treated with different agents. * p <0.05, ** p <0.01 and *** p < 0.001 were calculated by t test, referring to the statistically significant difference as compared to respective group.
Figure Legend Snippet: Analysis of the expression levels of gp91-phox and SOD1. ( A and B ) The expression levels of gp91-phox and SOD1 analyzed by Western Blot in LSCs and Non-LSCs treated with different agents. ( C – G ) Semi-quantification of the expression levels of gp91-phox, SOD1, CD38, DEPTOR, and IFITM3 detected by RT-qPCR in LSCs and Non-LSCs treated with different agents. * p <0.05, ** p <0.01 and *** p < 0.001 were calculated by t test, referring to the statistically significant difference as compared to respective group.

Techniques Used: Expressing, Western Blot, Quantitative RT-PCR


Structured Review

Proteintech gp91 phox
Effects of aging on mitochondrial stress and the redox state. (A) Mitochondrial ultrastructure of myocytes detected by electron microscopy; scale bar, 0.5 µm. (B) Representative immunoblots and (C) semi-quantitation of PGC-1α. (D) ROS levels detected by DHE fluorescence staining and (E) corresponding semi-quantification; scale bar, 50 µm. (F) Representative immunoblots of <t>p47-phox,</t> <t>gp91-phox,</t> CuZn- and Mn-SOD. (G) Semi-quantification of p47-phox and gp91-phox. (H) Semi-quantification of CuZn-SOD and Mn-SOD. Data are presented as the means ± standard error of the mean from each group; n=8 per group. *P<0.05, **P<0.01 vs. young group. DHE, dihydroethidium; NADPH, nicotinamide adenine dinucleotide phosphate; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α; ROS, reactive oxygen species; SOD, superoxide dismutase.
Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Age-related changes in mineralocorticoid receptors in rat hearts"

Article Title: Age-related changes in mineralocorticoid receptors in rat hearts

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2020.11260

Effects of aging on mitochondrial stress and the redox state. (A) Mitochondrial ultrastructure of myocytes detected by electron microscopy; scale bar, 0.5 µm. (B) Representative immunoblots and (C) semi-quantitation of PGC-1α. (D) ROS levels detected by DHE fluorescence staining and (E) corresponding semi-quantification; scale bar, 50 µm. (F) Representative immunoblots of p47-phox, gp91-phox, CuZn- and Mn-SOD. (G) Semi-quantification of p47-phox and gp91-phox. (H) Semi-quantification of CuZn-SOD and Mn-SOD. Data are presented as the means ± standard error of the mean from each group; n=8 per group. *P<0.05, **P<0.01 vs. young group. DHE, dihydroethidium; NADPH, nicotinamide adenine dinucleotide phosphate; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α; ROS, reactive oxygen species; SOD, superoxide dismutase.
Figure Legend Snippet: Effects of aging on mitochondrial stress and the redox state. (A) Mitochondrial ultrastructure of myocytes detected by electron microscopy; scale bar, 0.5 µm. (B) Representative immunoblots and (C) semi-quantitation of PGC-1α. (D) ROS levels detected by DHE fluorescence staining and (E) corresponding semi-quantification; scale bar, 50 µm. (F) Representative immunoblots of p47-phox, gp91-phox, CuZn- and Mn-SOD. (G) Semi-quantification of p47-phox and gp91-phox. (H) Semi-quantification of CuZn-SOD and Mn-SOD. Data are presented as the means ± standard error of the mean from each group; n=8 per group. *P<0.05, **P<0.01 vs. young group. DHE, dihydroethidium; NADPH, nicotinamide adenine dinucleotide phosphate; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α; ROS, reactive oxygen species; SOD, superoxide dismutase.

Techniques Used: Electron Microscopy, Western Blot, Quantitation Assay, Fluorescence, Staining

MR antagonism suppresses H 2 O 2 -induced cardiac aging and mitochondrial dysfunction. (A) Immunoblotting and semi-quantification of p16, p21, p53 and NADP subunits, p47-phox, p67-phox and gp91-phox expression in H9C2 cells. Protein expression levels were normalized to GAPDH. (B) ROS levels detected by DHE staining in H9C2 cells following different treatments (magnification, ×100; scale bar, 100 µm). (C) SOD-1, SOD-2 and PGC-1α protein expression levels detected by western blotting. Data are representative of three experiments, n=3. **P<0.01 and ***P<0.001 vs. control group; # P<0.05 vs. H 2 O 2 group. DHE, dihydroethidium; MR, mineralocorticoid receptors; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α; ROS, reactive oxygen species; SOD, superoxide dismutase; Eple, eplerenone.
Figure Legend Snippet: MR antagonism suppresses H 2 O 2 -induced cardiac aging and mitochondrial dysfunction. (A) Immunoblotting and semi-quantification of p16, p21, p53 and NADP subunits, p47-phox, p67-phox and gp91-phox expression in H9C2 cells. Protein expression levels were normalized to GAPDH. (B) ROS levels detected by DHE staining in H9C2 cells following different treatments (magnification, ×100; scale bar, 100 µm). (C) SOD-1, SOD-2 and PGC-1α protein expression levels detected by western blotting. Data are representative of three experiments, n=3. **P<0.01 and ***P<0.001 vs. control group; # P<0.05 vs. H 2 O 2 group. DHE, dihydroethidium; MR, mineralocorticoid receptors; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α; ROS, reactive oxygen species; SOD, superoxide dismutase; Eple, eplerenone.

Techniques Used: Western Blot, Expressing, Staining


Structured Review

Proteintech gp91 phox
Effect of gestational hypoxia compared to normoxia on protein expression in the oviducts of adult female rats
Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Chronic fetal hypoxia disrupts the peri‐conceptual environment in next‐generation adult female rats"

Article Title: Chronic fetal hypoxia disrupts the peri‐conceptual environment in next‐generation adult female rats

Journal: The Journal of Physiology

doi: 10.1113/JP277431

Effect of gestational hypoxia compared to normoxia on protein expression in the oviducts of adult female rats
Figure Legend Snippet: Effect of gestational hypoxia compared to normoxia on protein expression in the oviducts of adult female rats

Techniques Used: Expressing


Structured Review

Proteintech gp91 phox
Oxidative stress markers . The effect of in utero protein restriction and accelerated postnatal growth upon protein expression of (A) NF-κB, (B) markers of oxidative stress (XO, <t>Gp91</t> <t>phox</t> , P67 phox and cytochrome c ), (C) mRNA expression of Gp91 phox , P22 phox , P67 phox and Xo , (D) ETC complex activity, (E) citrate synthase (CS) activity and (F) Cox1 mRNA expression in vastus lateralis skeletal muscle in 12-month-old male rats. Results in A and B are shown as a percentage of the total amounts in control rats. Results are expressed as mean±s.e.m. * q <0.05 and ** P <0.01, *** P <0.001 (control versus recuperated). C, control; CS, citrate synthase; R, recuperated. n =6 per group for protein expression; n =8 per group for mRNA expression analysis; n =10 per group for ETC complex activity analysis.
Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Poor maternal nutrition and accelerated postnatal growth induces an accelerated aging phenotype and oxidative stress in skeletal muscle of male rats"

Article Title: Poor maternal nutrition and accelerated postnatal growth induces an accelerated aging phenotype and oxidative stress in skeletal muscle of male rats

Journal: Disease Models & Mechanisms

doi: 10.1242/dmm.026591

Oxidative stress markers . The effect of in utero protein restriction and accelerated postnatal growth upon protein expression of (A) NF-κB, (B) markers of oxidative stress (XO, Gp91 phox , P67 phox and cytochrome c ), (C) mRNA expression of Gp91 phox , P22 phox , P67 phox and Xo , (D) ETC complex activity, (E) citrate synthase (CS) activity and (F) Cox1 mRNA expression in vastus lateralis skeletal muscle in 12-month-old male rats. Results in A and B are shown as a percentage of the total amounts in control rats. Results are expressed as mean±s.e.m. * q <0.05 and ** P <0.01, *** P <0.001 (control versus recuperated). C, control; CS, citrate synthase; R, recuperated. n =6 per group for protein expression; n =8 per group for mRNA expression analysis; n =10 per group for ETC complex activity analysis.
Figure Legend Snippet: Oxidative stress markers . The effect of in utero protein restriction and accelerated postnatal growth upon protein expression of (A) NF-κB, (B) markers of oxidative stress (XO, Gp91 phox , P67 phox and cytochrome c ), (C) mRNA expression of Gp91 phox , P22 phox , P67 phox and Xo , (D) ETC complex activity, (E) citrate synthase (CS) activity and (F) Cox1 mRNA expression in vastus lateralis skeletal muscle in 12-month-old male rats. Results in A and B are shown as a percentage of the total amounts in control rats. Results are expressed as mean±s.e.m. * q <0.05 and ** P <0.01, *** P <0.001 (control versus recuperated). C, control; CS, citrate synthase; R, recuperated. n =6 per group for protein expression; n =8 per group for mRNA expression analysis; n =10 per group for ETC complex activity analysis.

Techniques Used: In Utero, Expressing, Activity Assay

Correlations of NF-κB protein expression with markers of oxidative stress, antioxidant enzymes and markers of inflammation . The effect of in utero protein restriction and accelerated postnatal growth upon correlations of protein expression of NF-κB versus (A) Gp91 phox , (B) XO, (C) MnSOD, (D) CuZnSOD, (E) catalase, (F) HO1 and (G) IL1β in vastus lateralis skeletal muscle of 12-month-old male rats. P -values are shown in the graphs (NF-κB versus antioxidants). Statistics were calculated using a Student's t -test (two-tailed). Results are expressed as mean±s.e.m. n =6 per group. IDV, integrated density value.
Figure Legend Snippet: Correlations of NF-κB protein expression with markers of oxidative stress, antioxidant enzymes and markers of inflammation . The effect of in utero protein restriction and accelerated postnatal growth upon correlations of protein expression of NF-κB versus (A) Gp91 phox , (B) XO, (C) MnSOD, (D) CuZnSOD, (E) catalase, (F) HO1 and (G) IL1β in vastus lateralis skeletal muscle of 12-month-old male rats. P -values are shown in the graphs (NF-κB versus antioxidants). Statistics were calculated using a Student's t -test (two-tailed). Results are expressed as mean±s.e.m. n =6 per group. IDV, integrated density value.

Techniques Used: Expressing, In Utero, Two Tailed Test

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    Proteintech anti gp91 phox
    a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress <t>GP91-PHOX</t> (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).
    Anti Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Proteintech gp91 phox antibodies
    a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress <t>GP91-PHOX</t> (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).
    Gp91 Phox Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gp91 phox antibodies/product/Proteintech
    Average 86 stars, based on 1 article reviews
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    94
    Proteintech gp91 phox
    Treatment of LXA4 inhibits oxidative stress in corpus cavernous of DMED rats. ( a ) Representative Western blot results for the p22 <t>phox</t> , p47 phox , <t>gp91</t> phox , p40 phox , and p67 phox in rats of all the three groups. ( b ) Expressions of p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox with β-actin as the loading control in all the three groups were presented through bar graphs. ( c ) MDA levels determined by the ELISA method in all the three groups. ( d ) SOD activities determined by the ELISA method in all the three groups. Data are expressed as mean ± s.e.m. ( n control = 10, n DMED = 11, and n DMED + LXA4 = 11). * P < 0.05 and *** P < 0.001 when comparing two groups. DMED: diabetes mellitus-induced erectile dysfunction; LXA4: Lipoxin A4; ELISA: enzyme-linked immunosorbent assay; SOD: superoxide dismutase; MDA: malondialdehyde; s.e.m.: standard error of the mean.
    Gp91 Phox, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress GP91-PHOX (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).

    Journal: Nature

    Article Title: Mitochondrial complex I activity in microglia sustains neuroinflammation

    doi: 10.1038/s41586-024-07167-9

    Figure Lengend Snippet: a , Heatmap of the log2-transformed counts of the top 25 differentially expressed genes (rows) involved in glycolysis and oxidative phosphorylation obtained from 22,148 cells (Ctrl = 6,205; A-EAE = 3,648; C-EAE = 12,295). Red text indicates genes of mitochondrial complex I (CI), III (CIII), and IV (CIV). b , c , UMAP plots of the whole scRNAseq dataset coloured by ( b ) stage of disease and ( c ) average expression of mitochondrial CI and CII genes (quantification is shown in the bar plots). d , Confocal images and quantification of the number of microglia (RFP + YFP + ) and infiltrating (RFP - YFP + ) myeloid cells expressing SPP1 and the marker of oxidative stress GP91-PHOX (n = 2, 3, 3 replicates per group; mean ± SEM; relevant statistical comparisons are shown with *P < 0.05, **P < 0.01, comparing total number of cells, and ##P < 0.01, comparing total number of GP91-PHOX + cells, two-way ANOVA; Fisher’s LSD).

    Article Snippet: The following primary antibodies were used (diluted in 1× PBS + 1% NGS or NDS and kept overnight at 4 °C): anti-GFP (chicken, 1:1,000, Abcam), anti-IBA1 (rabbit, 1:500, FUJIFILM Wako; or goat, 1:500, Novus biologicals), anti-CD3 (rat, 1:500, BD bioscience), anti-CD19 (rabbit, 1:500, Abcam), anti-GFAP (mouse, 1:400, Sigma-Aldrich) anti-CX3CR1 (rabbit, 1:300, Novus biologicals), anti-NEUN (mouse, 1:400, Sigma-Aldrich), anti-OLIG2 (goat, 1:500, Novus biologicals), anti-SPP1 (goat, 1:500, R&D Systems), anti-GP91-PHOX (rabbit, 1:500, Proteintech; or mouse, 1:200, BD bioscience) anti-CASPASE3 (mouse, 1:600, Novus biologicals), anti-NDUFS4 (mouse, 1:100, Abcam; or rabbit, 1:500, Novus biologicals), anti-amyloid precursor protein-APP (mouse, 1:200, Sigma-Aldrich), anti-neurofilament heavy polypeptide-NHP (rabbit, 1:500, Abcam).

    Techniques: Transformation Assay, Expressing, Marker

    a , Seahorse metabolic flux analysis of primary microglia derived from wild-type (WT) and Nd6 mice under basal conditions and after stimulation with LPS and IFNγ. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. Differences in basal respiration, mitochondrial (mt) ATP production and maximal respiration are reported. b , Quantification of mtROS production in LPS + IFNγ-stimulated (pro-inflammatory) primary WT and Nd6 microglia after RET induction (RET + ). From left to right, n = 18, 18, 17, 18, 18 and 18 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. c , Quantification of neuronal neurite length after co-culture with RET + pro-inflammatory primary WT and Nd6 microglia. From left to right, n = 11, 5, 6, 12, 12, 12 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. d , EAE scores of WT and Nd6 mice up to 30 days after immunization. n = 17 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. e , f , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 13,614 cells (7,501 (WT) and 6,113 (Nd6)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( e ) and EAE ( f ) mice (30 days after immunization). g , Selected hMG-like and DAM genes in cluster 0 and 1 DAMs. h , The mitochondrial membrane potential (Δ ψ m ) in ex vivo FACS-isolated CD45 + CD11b + cells. From left to right, n = 4, 3, 4 and 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Fisher’s LSD test. i , j Representative images and quantifications of perilesional microglial branching ( i ; n = 12 replicates per group) and IBA1 + SPP1 + GP91-PHOX + cells in WT and Nd6 EAE mice ( j ; n = 4 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. For i and j , scale bars, 30 μm. For d , i and j , data are mean ± s.e.m. The violin plots in a – c , and h show the median and quartiles. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

    Journal: Nature

    Article Title: Mitochondrial complex I activity in microglia sustains neuroinflammation

    doi: 10.1038/s41586-024-07167-9

    Figure Lengend Snippet: a , Seahorse metabolic flux analysis of primary microglia derived from wild-type (WT) and Nd6 mice under basal conditions and after stimulation with LPS and IFNγ. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. Differences in basal respiration, mitochondrial (mt) ATP production and maximal respiration are reported. b , Quantification of mtROS production in LPS + IFNγ-stimulated (pro-inflammatory) primary WT and Nd6 microglia after RET induction (RET + ). From left to right, n = 18, 18, 17, 18, 18 and 18 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. c , Quantification of neuronal neurite length after co-culture with RET + pro-inflammatory primary WT and Nd6 microglia. From left to right, n = 11, 5, 6, 12, 12, 12 replicates per group. Statistical analysis was performed using two-way ANOVA with Fisher’s LSD test. d , EAE scores of WT and Nd6 mice up to 30 days after immunization. n = 17 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. e , f , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 13,614 cells (7,501 (WT) and 6,113 (Nd6)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( e ) and EAE ( f ) mice (30 days after immunization). g , Selected hMG-like and DAM genes in cluster 0 and 1 DAMs. h , The mitochondrial membrane potential (Δ ψ m ) in ex vivo FACS-isolated CD45 + CD11b + cells. From left to right, n = 4, 3, 4 and 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Fisher’s LSD test. i , j Representative images and quantifications of perilesional microglial branching ( i ; n = 12 replicates per group) and IBA1 + SPP1 + GP91-PHOX + cells in WT and Nd6 EAE mice ( j ; n = 4 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. For i and j , scale bars, 30 μm. For d , i and j , data are mean ± s.e.m. The violin plots in a – c , and h show the median and quartiles. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

    Article Snippet: The following primary antibodies were used (diluted in 1× PBS + 1% NGS or NDS and kept overnight at 4 °C): anti-GFP (chicken, 1:1,000, Abcam), anti-IBA1 (rabbit, 1:500, FUJIFILM Wako; or goat, 1:500, Novus biologicals), anti-CD3 (rat, 1:500, BD bioscience), anti-CD19 (rabbit, 1:500, Abcam), anti-GFAP (mouse, 1:400, Sigma-Aldrich) anti-CX3CR1 (rabbit, 1:300, Novus biologicals), anti-NEUN (mouse, 1:400, Sigma-Aldrich), anti-OLIG2 (goat, 1:500, Novus biologicals), anti-SPP1 (goat, 1:500, R&D Systems), anti-GP91-PHOX (rabbit, 1:500, Proteintech; or mouse, 1:200, BD bioscience) anti-CASPASE3 (mouse, 1:600, Novus biologicals), anti-NDUFS4 (mouse, 1:100, Abcam; or rabbit, 1:500, Novus biologicals), anti-amyloid precursor protein-APP (mouse, 1:200, Sigma-Aldrich), anti-neurofilament heavy polypeptide-NHP (rabbit, 1:500, Abcam).

    Techniques: Derivative Assay, Co-Culture Assay, Membrane, Ex Vivo, Isolation, Two Tailed Test

    a , EAE scores of Ndufs4- WT and Ndufs4 -KO mice. n = 10 (WT) and 12 (KO) mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. b , c , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 10,666 cells (4,180 ( Ndufs4 WT) and 6,486 ( Ndufs4 KO)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( b ) and EAE ( c ) mice (50 days after immunization). d , Selected hMG-like and DAM genes in cluster 2 and 6 DAMs. e , Suspension mass cytometry (CyTOF) analysis of immune cell types at 50 days after immunization obtained from 51,177 cells (23,467 ( Ndufs4 WT); 27,710 ( Ndufs4 KO)). AA, alternatively activated; pro-inflam., pro-inflammatory. f , g , Quantification of CX3CR1 + SPP1 + ( f ; n = 5 replicates per group) and CASPASE3 + IBA1 + ( g ; n = 4 replicates per group) cells in EAE. Statistical analysis was performed using two-tailed unpaired t -tests. h – j , In vivo quantification of perilesional microglial branching ( h ; n = 12 (WT) and 11 (KO) replicates per group; two-tailed unpaired t -test), GP91-PHOX expression in the EAE spinal cords ( i ; n = 5 (WT) and 6 (KO) replicates per group; two-tailed Mann–Whitney U -test) and IBA1 + SPP1 + GP91-PHOX + cells ( j ; n = 4 replicates per group; two-tailed unpaired t -test). Scale bars, 7 μm ( h ) and 400 μm ( i ). k , l , Representative images and quantification of axonal loss ( k ; n = 5 (WT) and 6 (KO) replicates per group) and axonal degeneration ( l ; n = 5 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. APP, amyloid precursor protein; NHP, neurofilament heavy polypeptide. Insets: merged images. Scale bars, 400 μm ( k ) and 50 μm ( l ). m , EAE scores of mice treated with metformin, DMM, DMM + metformin versus saline controls. n = 13 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction; # P < 0.05 comparing DMM + metformin versus saline. n , CyTOF analysis of immune cell types at 30 days after immunization obtained from 159,110 cells (32,793 (metformin), 40,864 (DMM), 44,143 (DMM + metformin) and 41,310 (saline)). o , Quantification of CX3CR1 + SPP1 + NDUFS4 + cells, oxidative stress, axonal loss and axonal degeneration in EAE mice. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. For a , f – m and o , data are mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, *** P < 0.0001.

    Journal: Nature

    Article Title: Mitochondrial complex I activity in microglia sustains neuroinflammation

    doi: 10.1038/s41586-024-07167-9

    Figure Lengend Snippet: a , EAE scores of Ndufs4- WT and Ndufs4 -KO mice. n = 10 (WT) and 12 (KO) mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction. b , c , scRNA-seq UMAP plots with each cell coloured according to the genotype, obtained from 10,666 cells (4,180 ( Ndufs4 WT) and 6,486 ( Ndufs4 KO)). Superimposed cluster numbers and the corresponding fraction of cells are shown for control ( b ) and EAE ( c ) mice (50 days after immunization). d , Selected hMG-like and DAM genes in cluster 2 and 6 DAMs. e , Suspension mass cytometry (CyTOF) analysis of immune cell types at 50 days after immunization obtained from 51,177 cells (23,467 ( Ndufs4 WT); 27,710 ( Ndufs4 KO)). AA, alternatively activated; pro-inflam., pro-inflammatory. f , g , Quantification of CX3CR1 + SPP1 + ( f ; n = 5 replicates per group) and CASPASE3 + IBA1 + ( g ; n = 4 replicates per group) cells in EAE. Statistical analysis was performed using two-tailed unpaired t -tests. h – j , In vivo quantification of perilesional microglial branching ( h ; n = 12 (WT) and 11 (KO) replicates per group; two-tailed unpaired t -test), GP91-PHOX expression in the EAE spinal cords ( i ; n = 5 (WT) and 6 (KO) replicates per group; two-tailed Mann–Whitney U -test) and IBA1 + SPP1 + GP91-PHOX + cells ( j ; n = 4 replicates per group; two-tailed unpaired t -test). Scale bars, 7 μm ( h ) and 400 μm ( i ). k , l , Representative images and quantification of axonal loss ( k ; n = 5 (WT) and 6 (KO) replicates per group) and axonal degeneration ( l ; n = 5 replicates per group). Statistical analysis was performed using two-tailed unpaired t -tests. APP, amyloid precursor protein; NHP, neurofilament heavy polypeptide. Insets: merged images. Scale bars, 400 μm ( k ) and 50 μm ( l ). m , EAE scores of mice treated with metformin, DMM, DMM + metformin versus saline controls. n = 13 mice per group. Statistical analysis was performed using two-way ANOVA with Bonferroni correction; # P < 0.05 comparing DMM + metformin versus saline. n , CyTOF analysis of immune cell types at 30 days after immunization obtained from 159,110 cells (32,793 (metformin), 40,864 (DMM), 44,143 (DMM + metformin) and 41,310 (saline)). o , Quantification of CX3CR1 + SPP1 + NDUFS4 + cells, oxidative stress, axonal loss and axonal degeneration in EAE mice. n = 4 replicates per group. Statistical analysis was performed using one-way ANOVA with Tukey test. For a , f – m and o , data are mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, *** P < 0.0001.

    Article Snippet: The following primary antibodies were used (diluted in 1× PBS + 1% NGS or NDS and kept overnight at 4 °C): anti-GFP (chicken, 1:1,000, Abcam), anti-IBA1 (rabbit, 1:500, FUJIFILM Wako; or goat, 1:500, Novus biologicals), anti-CD3 (rat, 1:500, BD bioscience), anti-CD19 (rabbit, 1:500, Abcam), anti-GFAP (mouse, 1:400, Sigma-Aldrich) anti-CX3CR1 (rabbit, 1:300, Novus biologicals), anti-NEUN (mouse, 1:400, Sigma-Aldrich), anti-OLIG2 (goat, 1:500, Novus biologicals), anti-SPP1 (goat, 1:500, R&D Systems), anti-GP91-PHOX (rabbit, 1:500, Proteintech; or mouse, 1:200, BD bioscience) anti-CASPASE3 (mouse, 1:600, Novus biologicals), anti-NDUFS4 (mouse, 1:100, Abcam; or rabbit, 1:500, Novus biologicals), anti-amyloid precursor protein-APP (mouse, 1:200, Sigma-Aldrich), anti-neurofilament heavy polypeptide-NHP (rabbit, 1:500, Abcam).

    Techniques: Suspension, Mass Cytometry, Two Tailed Test, In Vivo, Expressing, MANN-WHITNEY, Saline

    a , Cytotoxicity assay and mtROS production of mouse BV2 microglia treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 7, 3, 3, 3, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (first column, top-bottom), n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 37, 11, 14, 11, 11, 10, 11, 6, 6, 6, 3, 11, 11 replicates per group (first column, top-bottom), n = 46, 11, 14, 11, 11, 18, 17, 5, 6, 6, 3, 18, 18 replicates per group (second column, top-bottom), n = 38, 20, 16, 20, 20, 18, 18, 8, 8, 8, 4, 18, 18 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. b , Cytotoxicity assay and mtROS production of human induced microglia (hiMG) treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (first column, top-bottom), n = 7, 4, 3, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 27, 10, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (first column, top-bottom), n = 32, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (second column, top-bottom), n = 31, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 5 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. c , EAE scores of mice treated with 4-octyl itaconate (4-OI) vs saline controls (n = 6 mice per group; mean ± SEM; two-way ANOVA; Bonferroni). d , e , In vivo testing of dimethyl malonate (DMM) and disodium malonate (DSM) in EAE. ( d ) EAE mice receiving daily intraperitoneal (IP) injections of DMM (160 mg/kg), DSM (160 mg/kg), or saline at 7 days from disease onset. Malonate levels in the peripheral blood (at 30 min from injection, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). ***P < 0.001. ( e ) EAE mice receiving DMM (1.5%), DSM (1.5%), or saline dissolved in their drinking water (at 7 days from disease onset). Malonate levels in the peripheral blood during treatment (daily, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). *P < 0.05, **P < 0.01. f , UMAP plot from the CyTOF data at 30 dpi (see also Fig. ) coloured by cell type found in EAE mice treated with CII and CI inhibitors. Numbers of clusters are shown in superimposition. A.A.: alternatively activated. g , Fraction of cell clusters in EAE mice treated with metformin, DMM, DMM+metformin, vs saline-treated controls at 30 dpi. h , i , Expression of ( h ) CII and ( i ) CI in the CyTOF clusters identified as hMG-like cells and DAM (median and quartiles from 159,110 cells; *P < 0.01, **P < 0.0001; unpaired t-test, two-tailed). j , Representative confocal images and quantification of the expression of the marker of oxidative stress GP91-PHOX in microglia (IBA1 + ), astrocytes (GFAP + ), oligodendrocytes (OLIG2 + ), and neurons (NEUN + ) in the spinal cord of treated EAE mice (n = 4 replicates per group; mean ± SEM; *P < 0.05; one-way ANOVA; Tukey). Abbreviations: rotenone (rot.), metformin (met.), 4-octyl itaconate (4-OI), dimethyl malonate (DMM), disodium malonate (DSM).

    Journal: Nature

    Article Title: Mitochondrial complex I activity in microglia sustains neuroinflammation

    doi: 10.1038/s41586-024-07167-9

    Figure Lengend Snippet: a , Cytotoxicity assay and mtROS production of mouse BV2 microglia treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 7, 3, 3, 3, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (first column, top-bottom), n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 15 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 37, 11, 14, 11, 11, 10, 11, 6, 6, 6, 3, 11, 11 replicates per group (first column, top-bottom), n = 46, 11, 14, 11, 11, 18, 17, 5, 6, 6, 3, 18, 18 replicates per group (second column, top-bottom), n = 38, 20, 16, 20, 20, 18, 18, 8, 8, 8, 4, 18, 18 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. b , Cytotoxicity assay and mtROS production of human induced microglia (hiMG) treated with selected CI and/or CII inhibitors in vitro. Cytotoxicity assay data are expressed as % of death control from n = 8, 4, 4, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (first column, top-bottom), n = 7, 4, 3, 4, 4, 4, 4, 4, 4, 4, 4, 16 replicates per group (second column, top-bottom); *P < 0.05 vs death control; one-way ANOVA; Tukey). MtROS data is normalized on RET and shown as % increase or decrease vs RET mtROS baseline (assessed via MitoSOX) from n = 27, 10, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (first column, top-bottom), n = 32, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 6 replicates per group (second column, top-bottom), n = 31, 26, 10, 10, 10, 6, 6, 8, 8, 8, 8, 6, 5 replicates per group (third column, top-bottom); *P < 0.05, **P < 0.01 vs untreated RET + pro-inflammatory microglia; unpaired t-test, two-tailed. c , EAE scores of mice treated with 4-octyl itaconate (4-OI) vs saline controls (n = 6 mice per group; mean ± SEM; two-way ANOVA; Bonferroni). d , e , In vivo testing of dimethyl malonate (DMM) and disodium malonate (DSM) in EAE. ( d ) EAE mice receiving daily intraperitoneal (IP) injections of DMM (160 mg/kg), DSM (160 mg/kg), or saline at 7 days from disease onset. Malonate levels in the peripheral blood (at 30 min from injection, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). ***P < 0.001. ( e ) EAE mice receiving DMM (1.5%), DSM (1.5%), or saline dissolved in their drinking water (at 7 days from disease onset). Malonate levels in the peripheral blood during treatment (daily, left; n = 5 replicates per group; mean ± SEM; one-way ANOVA; Tukey) and in the CNS (at sacrifice, right; n = 2 replicates per group; mean). *P < 0.05, **P < 0.01. f , UMAP plot from the CyTOF data at 30 dpi (see also Fig. ) coloured by cell type found in EAE mice treated with CII and CI inhibitors. Numbers of clusters are shown in superimposition. A.A.: alternatively activated. g , Fraction of cell clusters in EAE mice treated with metformin, DMM, DMM+metformin, vs saline-treated controls at 30 dpi. h , i , Expression of ( h ) CII and ( i ) CI in the CyTOF clusters identified as hMG-like cells and DAM (median and quartiles from 159,110 cells; *P < 0.01, **P < 0.0001; unpaired t-test, two-tailed). j , Representative confocal images and quantification of the expression of the marker of oxidative stress GP91-PHOX in microglia (IBA1 + ), astrocytes (GFAP + ), oligodendrocytes (OLIG2 + ), and neurons (NEUN + ) in the spinal cord of treated EAE mice (n = 4 replicates per group; mean ± SEM; *P < 0.05; one-way ANOVA; Tukey). Abbreviations: rotenone (rot.), metformin (met.), 4-octyl itaconate (4-OI), dimethyl malonate (DMM), disodium malonate (DSM).

    Article Snippet: The following primary antibodies were used (diluted in 1× PBS + 1% NGS or NDS and kept overnight at 4 °C): anti-GFP (chicken, 1:1,000, Abcam), anti-IBA1 (rabbit, 1:500, FUJIFILM Wako; or goat, 1:500, Novus biologicals), anti-CD3 (rat, 1:500, BD bioscience), anti-CD19 (rabbit, 1:500, Abcam), anti-GFAP (mouse, 1:400, Sigma-Aldrich) anti-CX3CR1 (rabbit, 1:300, Novus biologicals), anti-NEUN (mouse, 1:400, Sigma-Aldrich), anti-OLIG2 (goat, 1:500, Novus biologicals), anti-SPP1 (goat, 1:500, R&D Systems), anti-GP91-PHOX (rabbit, 1:500, Proteintech; or mouse, 1:200, BD bioscience) anti-CASPASE3 (mouse, 1:600, Novus biologicals), anti-NDUFS4 (mouse, 1:100, Abcam; or rabbit, 1:500, Novus biologicals), anti-amyloid precursor protein-APP (mouse, 1:200, Sigma-Aldrich), anti-neurofilament heavy polypeptide-NHP (rabbit, 1:500, Abcam).

    Techniques: Cytotoxicity Assay, In Vitro, Two Tailed Test, Saline, In Vivo, Injection, Expressing, Marker

    Treatment of LXA4 inhibits oxidative stress in corpus cavernous of DMED rats. ( a ) Representative Western blot results for the p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox in rats of all the three groups. ( b ) Expressions of p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox with β-actin as the loading control in all the three groups were presented through bar graphs. ( c ) MDA levels determined by the ELISA method in all the three groups. ( d ) SOD activities determined by the ELISA method in all the three groups. Data are expressed as mean ± s.e.m. ( n control = 10, n DMED = 11, and n DMED + LXA4 = 11). * P < 0.05 and *** P < 0.001 when comparing two groups. DMED: diabetes mellitus-induced erectile dysfunction; LXA4: Lipoxin A4; ELISA: enzyme-linked immunosorbent assay; SOD: superoxide dismutase; MDA: malondialdehyde; s.e.m.: standard error of the mean.

    Journal: Asian Journal of Andrology

    Article Title: Lipoxin A4 improves erectile dysfunction in rats with type I diabetes by inhibiting oxidative stress and corporal fibrosis

    doi: 10.4103/aja.aja_49_17

    Figure Lengend Snippet: Treatment of LXA4 inhibits oxidative stress in corpus cavernous of DMED rats. ( a ) Representative Western blot results for the p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox in rats of all the three groups. ( b ) Expressions of p22 phox , p47 phox , gp91 phox , p40 phox , and p67 phox with β-actin as the loading control in all the three groups were presented through bar graphs. ( c ) MDA levels determined by the ELISA method in all the three groups. ( d ) SOD activities determined by the ELISA method in all the three groups. Data are expressed as mean ± s.e.m. ( n control = 10, n DMED = 11, and n DMED + LXA4 = 11). * P < 0.05 and *** P < 0.001 when comparing two groups. DMED: diabetes mellitus-induced erectile dysfunction; LXA4: Lipoxin A4; ELISA: enzyme-linked immunosorbent assay; SOD: superoxide dismutase; MDA: malondialdehyde; s.e.m.: standard error of the mean.

    Article Snippet: Equal amounts of protein samples (30 μg per lane) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes, followed by blocking in Tris-buffered saline-Tween (TBST) with 5% bovine serum albumin (BSA) for 1 h. Then, membranes were incubated with primary antibodies against: eNOS (1:1000; ab5589, Abcam, Cambridge, MA, USA), p-eNOS (Ser1177; 1:1000; 9571; Cell Signaling Technology, Danvers, MA, USA), neuronal nitric oxide synthase (nNOS; 1:1000; ab76067; Abcam), p22 phox (1:500; sc-271968, Santa Cruz, Dallas, Texas, USA), p47 phox (1:1000; D154042, Sangon Biotech, Shanghai, China), gp91 phox (1:500, 19013-1-AP, Proteintech); p40 phox (1:500; D154073, Sangon Biotech), p67 phox (1:1000; 15551-1-AP, Proteintech), α-smooth muscle actin (α-SMA; 1:1000; ab5694; Abcam), Collagen I (1:500; PB0981, Boster, Wuhan, China), Collagen IV (1:500; 19797-1-AP, Proteintech), TGF-β1 (1:1000; ab92486, Abcam), Smad2/3 (1:1000; 8685, Cell Signaling Technology), p-Smad2/3 (1:1000; 8828, Cell Signaling Technology), CTGF (1:1000; 23936-1-AP, Proteintech), and β-actin (1:1000, 20536-1-AP, Proteintech).

    Techniques: Western Blot, Enzyme-linked Immunosorbent Assay