Structured Review

Santa Cruz Biotechnology rabbit anti p22 phox
a. Schematic of experiment for disrupting ETS2 in primary monocytes and differentiating monocyte-derived macrophages under chronic inflammatory conditions. b. Macrophage cytokine secretion following ETS2 disruption. Heatmap shows log2 fold-change of cytokine concentrations in the supernatants of ETS2 -edited macrophages relative to unedited macrophages transfected with a non-targeting control gRNA-containing RNP (NTC). n=9, Wilcoxon matched-pairs test, two-tailed. c. Histogram depicting phagocytosis of fluorescently-labelled zymosan particles by ETS2 -edited and unedited macrophages (left). Data representative of one of seven donors. Phagocytosis index in ETS2 -edited and unedited macrophages (calculated as product of proportion and mean fluorescence intensity of phagocytosing cells; right). Plot depicts log2 fold-change in phagocytosis index for ETS2 -edited macrophages relative to unedited cells (Wilcoxon signed-rank test, two-tailed; data represent mean±SEM). d. Production of ROS by ETS2 -edited and unedited inflammatory macrophages (measured in relative light units; left). Data representative of one of six donors. Western blot for gp91 <t>phox</t> , <t>p22</t> phox , and EROS expression in ETS2 -edited and unedited macrophages (right). Data representative of one of three donors. e. Differentially-expressed genes in ETS2 -edited versus unedited inflammatory macrophages (limma with voom transformation, n=8). f. Gene set enrichment analysis (fGSEA) of differentially-expressed genes between ETS2 -edited and unedited inflammatory macrophages. Results of selected Gene Ontology Biological Pathways shown. Dot size represents P -value and colour denotes normalised enrichment score (NES). g. Enrichment of differentially-expressed genes following deletion of the disease-associated chr21q22 locus (upregulated genes, top; downregulated genes, bottom) in ETS2 -edited versus unedited macrophages. * P < 0.05, ** P < 0.01.
Rabbit Anti P22 Phox, supplied by Santa Cruz Biotechnology, 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 "A disease-associated gene desert orchestrates macrophage inflammatory responses via ETS2"

Article Title: A disease-associated gene desert orchestrates macrophage inflammatory responses via ETS2

Journal: bioRxiv

doi: 10.1101/2023.05.05.539522

a. Schematic of experiment for disrupting ETS2 in primary monocytes and differentiating monocyte-derived macrophages under chronic inflammatory conditions. b. Macrophage cytokine secretion following ETS2 disruption. Heatmap shows log2 fold-change of cytokine concentrations in the supernatants of ETS2 -edited macrophages relative to unedited macrophages transfected with a non-targeting control gRNA-containing RNP (NTC). n=9, Wilcoxon matched-pairs test, two-tailed. c. Histogram depicting phagocytosis of fluorescently-labelled zymosan particles by ETS2 -edited and unedited macrophages (left). Data representative of one of seven donors. Phagocytosis index in ETS2 -edited and unedited macrophages (calculated as product of proportion and mean fluorescence intensity of phagocytosing cells; right). Plot depicts log2 fold-change in phagocytosis index for ETS2 -edited macrophages relative to unedited cells (Wilcoxon signed-rank test, two-tailed; data represent mean±SEM). d. Production of ROS by ETS2 -edited and unedited inflammatory macrophages (measured in relative light units; left). Data representative of one of six donors. Western blot for gp91 phox , p22 phox , and EROS expression in ETS2 -edited and unedited macrophages (right). Data representative of one of three donors. e. Differentially-expressed genes in ETS2 -edited versus unedited inflammatory macrophages (limma with voom transformation, n=8). f. Gene set enrichment analysis (fGSEA) of differentially-expressed genes between ETS2 -edited and unedited inflammatory macrophages. Results of selected Gene Ontology Biological Pathways shown. Dot size represents P -value and colour denotes normalised enrichment score (NES). g. Enrichment of differentially-expressed genes following deletion of the disease-associated chr21q22 locus (upregulated genes, top; downregulated genes, bottom) in ETS2 -edited versus unedited macrophages. * P < 0.05, ** P < 0.01.
Figure Legend Snippet: a. Schematic of experiment for disrupting ETS2 in primary monocytes and differentiating monocyte-derived macrophages under chronic inflammatory conditions. b. Macrophage cytokine secretion following ETS2 disruption. Heatmap shows log2 fold-change of cytokine concentrations in the supernatants of ETS2 -edited macrophages relative to unedited macrophages transfected with a non-targeting control gRNA-containing RNP (NTC). n=9, Wilcoxon matched-pairs test, two-tailed. c. Histogram depicting phagocytosis of fluorescently-labelled zymosan particles by ETS2 -edited and unedited macrophages (left). Data representative of one of seven donors. Phagocytosis index in ETS2 -edited and unedited macrophages (calculated as product of proportion and mean fluorescence intensity of phagocytosing cells; right). Plot depicts log2 fold-change in phagocytosis index for ETS2 -edited macrophages relative to unedited cells (Wilcoxon signed-rank test, two-tailed; data represent mean±SEM). d. Production of ROS by ETS2 -edited and unedited inflammatory macrophages (measured in relative light units; left). Data representative of one of six donors. Western blot for gp91 phox , p22 phox , and EROS expression in ETS2 -edited and unedited macrophages (right). Data representative of one of three donors. e. Differentially-expressed genes in ETS2 -edited versus unedited inflammatory macrophages (limma with voom transformation, n=8). f. Gene set enrichment analysis (fGSEA) of differentially-expressed genes between ETS2 -edited and unedited inflammatory macrophages. Results of selected Gene Ontology Biological Pathways shown. Dot size represents P -value and colour denotes normalised enrichment score (NES). g. Enrichment of differentially-expressed genes following deletion of the disease-associated chr21q22 locus (upregulated genes, top; downregulated genes, bottom) in ETS2 -edited versus unedited macrophages. * P < 0.05, ** P < 0.01.

Techniques Used: Derivative Assay, Transfection, Two Tailed Test, Fluorescence, Western Blot, Expressing, Transformation Assay


Structured Review

Santa Cruz Biotechnology rabbit polyclonal antibodies against p22 phox
Nox-4 (a) and <t>p22</t> <t>phox</t> (b) protein expressions in the liver. m/m : Misty; Veh: vehicle-treated db/db mice; K-100: Kangen-karyu 100 mg/kg body weight-treated db/db mice; K-200: Kangen-karyu 200 mg/kg body weight-treated db/db mice. Bars represent the means ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001 versus vehicle-treated db/db mouse values.
Rabbit Polyclonal Antibodies Against P22 Phox, supplied by Santa Cruz Biotechnology, 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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1) Product Images from "Evaluation of Effects of Chinese Prescription Kangen-karyu on Diabetes-Induced Alterations such as Oxidative Stress and Apoptosis in the Liver of Type 2 Diabetic db/db Mice"

Article Title: Evaluation of Effects of Chinese Prescription Kangen-karyu on Diabetes-Induced Alterations such as Oxidative Stress and Apoptosis in the Liver of Type 2 Diabetic db/db Mice

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2012/143489

Nox-4 (a) and p22 phox (b) protein expressions in the liver. m/m : Misty; Veh: vehicle-treated db/db mice; K-100: Kangen-karyu 100 mg/kg body weight-treated db/db mice; K-200: Kangen-karyu 200 mg/kg body weight-treated db/db mice. Bars represent the means ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001 versus vehicle-treated db/db mouse values.
Figure Legend Snippet: Nox-4 (a) and p22 phox (b) protein expressions in the liver. m/m : Misty; Veh: vehicle-treated db/db mice; K-100: Kangen-karyu 100 mg/kg body weight-treated db/db mice; K-200: Kangen-karyu 200 mg/kg body weight-treated db/db mice. Bars represent the means ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001 versus vehicle-treated db/db mouse values.

Techniques Used:


Structured Review

Santa Cruz Biotechnology rabbit p22 phox
( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to <t>p22</t> <t>phox</t> (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.
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1) Product Images from "Reactive Oxygen Species Regulate the Levels of Dual Oxidase (Duox1-2) in Human Neuroblastoma Cells"

Article Title: Reactive Oxygen Species Regulate the Levels of Dual Oxidase (Duox1-2) in Human Neuroblastoma Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0034405

( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.
Figure Legend Snippet: ( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.

Techniques Used: Transfection, Electroporation, Western Blot, Incubation, Reverse Transcription Polymerase Chain Reaction

( A ) Cells were incubated 18h in medium containing 0.2% FBS, loaded with 10µM DCHF-DA in the presence or absence of the intracellular calcium chelator, BAPTA-AM (10µM), and then stimulated with 15ng/ml of PDGF as described in . ROS levels were measured fluorimetrically at the time intervals indicated. Ca ++ -independent ROS were measured in presence of BAPTA-AM. Ca ++ -dependent ROS levels were derived from the assays performed in the presence or absence of BAPTA-AM. Total levels of ROS induced by PDGF were also measured in the absence (Total) or presence of the NADPH oxidase inhibitor AEBSF. Values are Mean +/- SEM of three independent experiments performed in triplicate. * p< 0.01 and ** p< 0.05 vs not stimulated; § p<0.01 vs the corresponding time point of Total (Ctr) curve. ( B ) Cells were transfected by electroporation with siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 24h after transfection cells were incubated in medium containing 0.2%FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. An aliquot of cell medium was collected and analyzed for H 2 O 2 levels as described in . The histograms show the mean +/- SEM values of three independent experiments. * p< 0.01 vs Ctr. ** p< 0.01 vs PDGF stimulated scramble.
Figure Legend Snippet: ( A ) Cells were incubated 18h in medium containing 0.2% FBS, loaded with 10µM DCHF-DA in the presence or absence of the intracellular calcium chelator, BAPTA-AM (10µM), and then stimulated with 15ng/ml of PDGF as described in . ROS levels were measured fluorimetrically at the time intervals indicated. Ca ++ -independent ROS were measured in presence of BAPTA-AM. Ca ++ -dependent ROS levels were derived from the assays performed in the presence or absence of BAPTA-AM. Total levels of ROS induced by PDGF were also measured in the absence (Total) or presence of the NADPH oxidase inhibitor AEBSF. Values are Mean +/- SEM of three independent experiments performed in triplicate. * p< 0.01 and ** p< 0.05 vs not stimulated; § p<0.01 vs the corresponding time point of Total (Ctr) curve. ( B ) Cells were transfected by electroporation with siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 24h after transfection cells were incubated in medium containing 0.2%FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. An aliquot of cell medium was collected and analyzed for H 2 O 2 levels as described in . The histograms show the mean +/- SEM values of three independent experiments. * p< 0.01 vs Ctr. ** p< 0.01 vs PDGF stimulated scramble.

Techniques Used: Incubation, Derivative Assay, Transfection, Electroporation


Structured Review

Santa Cruz Biotechnology rabbit p22 phox
The primer sequences of <t> p22 phox </t> , p47 phox , and GAPDH.
Rabbit P22 Phox, supplied by Santa Cruz Biotechnology, 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 "Pioglitazone Inhibits the Expressions of p22 phox and p47 phox in Rat Mesangial Cells In Vitro"

Article Title: Pioglitazone Inhibits the Expressions of p22 phox and p47 phox in Rat Mesangial Cells In Vitro

Journal: ISRN Endocrinology

doi: 10.1155/2014/601352

The primer sequences of  p22 phox  , p47 phox , and GAPDH.
Figure Legend Snippet: The primer sequences of p22 phox , p47 phox , and GAPDH.

Techniques Used:

Expressions of p22 phox and p47 phox mRNA in mesangial cells evaluated by RT-PCR. Gene expressions of p22 phox (a), p47 phox (b), and GAPDH (c) in mesangial cells. M: maker; 1: NG; 2: HG; 3: PIO 1; 4: PIO 2; 5: PIO 3. Expressions of p22 phox (d) and p47 phox (e) mRNA among different groups. The values (means ± SD) from six independent experiments are relative to the NG group, which is set as 1.0. * P < 0.01 versus NG group; # P < 0.05 versus HG group.
Figure Legend Snippet: Expressions of p22 phox and p47 phox mRNA in mesangial cells evaluated by RT-PCR. Gene expressions of p22 phox (a), p47 phox (b), and GAPDH (c) in mesangial cells. M: maker; 1: NG; 2: HG; 3: PIO 1; 4: PIO 2; 5: PIO 3. Expressions of p22 phox (d) and p47 phox (e) mRNA among different groups. The values (means ± SD) from six independent experiments are relative to the NG group, which is set as 1.0. * P < 0.01 versus NG group; # P < 0.05 versus HG group.

Techniques Used: Reverse Transcription Polymerase Chain Reaction

Effects of pioglitazone on protein expression of p22 phox and p47 phox in HG-treated mesangial cells measured by western blot. (a) Pioglitazone inhibited p22 phox expression in mesangial cells. (b) Pioglitazone inhibited p47 phox expression in mesangial cells. The values (means ± SD) from six independent experiments are relative to the control, which is set as 1.0. * P < 0.01 versus NG group; # P < 0.05 versus HG group.
Figure Legend Snippet: Effects of pioglitazone on protein expression of p22 phox and p47 phox in HG-treated mesangial cells measured by western blot. (a) Pioglitazone inhibited p22 phox expression in mesangial cells. (b) Pioglitazone inhibited p47 phox expression in mesangial cells. The values (means ± SD) from six independent experiments are relative to the control, which is set as 1.0. * P < 0.01 versus NG group; # P < 0.05 versus HG group.

Techniques Used: Expressing, Western Blot


Structured Review

Santa Cruz Biotechnology rabbit polyclonal anti p22 phox
Rabbit Polyclonal Anti P22 Phox, supplied by Santa Cruz Biotechnology, 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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Santa Cruz Biotechnology rabbit polyclonal antibodies against p22 phox
NADPH oxidase subunit <t>p22</t> <t>phox</t> expression. Mesangial cells were exposed to 5.6 mM (NG) or 25 mM (HG) D-glucose for 24 hours or 48 hours, or pretreated with 10 μ M rosiglitazone (RSG), or 10 μ M ciglitazone or 10 μ M troglitazone) and p22 phox was detected by immunoblotting. Pretreatment with three different PPAR γ agonists blocked HG-induced p22 phox expression ( n = 5, * P < .05 versus NG, ** P < .05 versus HG 24 hours or HG 48 hours).
Rabbit Polyclonal Antibodies Against P22 Phox, supplied by Santa Cruz Biotechnology, 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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1) Product Images from "Rosiglitazone Prevents High Glucose-Induced Vascular Endothelial Growth Factor and Collagen IV Expression in Cultured Mesangial Cells"

Article Title: Rosiglitazone Prevents High Glucose-Induced Vascular Endothelial Growth Factor and Collagen IV Expression in Cultured Mesangial Cells

Journal: Experimental Diabetes Research

doi: 10.1155/2009/910783

NADPH oxidase subunit p22 phox expression. Mesangial cells were exposed to 5.6 mM (NG) or 25 mM (HG) D-glucose for 24 hours or 48 hours, or pretreated with 10 μ M rosiglitazone (RSG), or 10 μ M ciglitazone or 10 μ M troglitazone) and p22 phox was detected by immunoblotting. Pretreatment with three different PPAR γ agonists blocked HG-induced p22 phox expression ( n = 5, * P < .05 versus NG, ** P < .05 versus HG 24 hours or HG 48 hours).
Figure Legend Snippet: NADPH oxidase subunit p22 phox expression. Mesangial cells were exposed to 5.6 mM (NG) or 25 mM (HG) D-glucose for 24 hours or 48 hours, or pretreated with 10 μ M rosiglitazone (RSG), or 10 μ M ciglitazone or 10 μ M troglitazone) and p22 phox was detected by immunoblotting. Pretreatment with three different PPAR γ agonists blocked HG-induced p22 phox expression ( n = 5, * P < .05 versus NG, ** P < .05 versus HG 24 hours or HG 48 hours).

Techniques Used: Expressing, Western Blot


Structured Review

Santa Cruz Biotechnology rabbit polyclonal anti p22 phox
(A) Expression of the NADPH oxidase subunits Nox4 and <t>p22</t> <t>phox</t> in the renal cortex of young and old WKY rats. GAPDH was used to control for loading differences. (B) The bar graph shows a summary of densitometric data. Values are normalized to the level of GAPDH expression in each condition and expressed as % of young rats. Each column represents the mean ± SEM of six independent immunoblots. Significantly different from values in young rats (*p < 0.05) using the Newman-Keuls test.
Rabbit Polyclonal Anti P22 Phox, supplied by Santa Cruz Biotechnology, 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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1) Product Images from "Aging increases oxidative stress and renal expression of oxidant and antioxidant enzymes that are associated with an increased trend in systolic blood pressure"

Article Title: Aging increases oxidative stress and renal expression of oxidant and antioxidant enzymes that are associated with an increased trend in systolic blood pressure

Journal: Oxidative Medicine and Cellular Longevity

doi:

(A) Expression of the NADPH oxidase subunits Nox4 and p22 phox in the renal cortex of young and old WKY rats. GAPDH was used to control for loading differences. (B) The bar graph shows a summary of densitometric data. Values are normalized to the level of GAPDH expression in each condition and expressed as % of young rats. Each column represents the mean ± SEM of six independent immunoblots. Significantly different from values in young rats (*p < 0.05) using the Newman-Keuls test.
Figure Legend Snippet: (A) Expression of the NADPH oxidase subunits Nox4 and p22 phox in the renal cortex of young and old WKY rats. GAPDH was used to control for loading differences. (B) The bar graph shows a summary of densitometric data. Values are normalized to the level of GAPDH expression in each condition and expressed as % of young rats. Each column represents the mean ± SEM of six independent immunoblots. Significantly different from values in young rats (*p < 0.05) using the Newman-Keuls test.

Techniques Used: Expressing, Western Blot


Structured Review

Santa Cruz Biotechnology rabbit antibodies against p22 phox
A. Representative western blots of all NADPH oxidase subunits from TA muscles of 18–19 day old WT and mdx mice. B. Pooled results from densitometry for the subunits showing mdx values normalized to WT (represented by the blue dotted line). The proteins gp91 <t>phox</t> , p67 phox and rac1 were all significantly increased in mdx muscle compared to WT, whereas <t>p22</t> phox and p47 phox were not different (n = 4 samples for each group).
Rabbit Antibodies Against P22 Phox, supplied by Santa Cruz Biotechnology, 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 "Skeletal Muscle NADPH Oxidase Is Increased and Triggers Stretch-Induced Damage in the mdx Mouse"

Article Title: Skeletal Muscle NADPH Oxidase Is Increased and Triggers Stretch-Induced Damage in the mdx Mouse

Journal: PLoS ONE

doi: 10.1371/journal.pone.0015354

A. Representative western blots of all NADPH oxidase subunits from TA muscles of 18–19 day old WT and mdx mice. B. Pooled results from densitometry for the subunits showing mdx values normalized to WT (represented by the blue dotted line). The proteins gp91 phox , p67 phox and rac1 were all significantly increased in mdx muscle compared to WT, whereas p22 phox and p47 phox were not different (n = 4 samples for each group).
Figure Legend Snippet: A. Representative western blots of all NADPH oxidase subunits from TA muscles of 18–19 day old WT and mdx mice. B. Pooled results from densitometry for the subunits showing mdx values normalized to WT (represented by the blue dotted line). The proteins gp91 phox , p67 phox and rac1 were all significantly increased in mdx muscle compared to WT, whereas p22 phox and p47 phox were not different (n = 4 samples for each group).

Techniques Used: Western Blot

Isolated mdx muscle fibers from the FDB muscle were immunostained using antibodies against the various NADPH oxidase subunits. Nuclei are stained by DAPI (blue). In the bottom right panel, p22 phox (green) was co-immunostained with caveolin-3 (red) to demonstrate sarcolemmal localization (yellow).
Figure Legend Snippet: Isolated mdx muscle fibers from the FDB muscle were immunostained using antibodies against the various NADPH oxidase subunits. Nuclei are stained by DAPI (blue). In the bottom right panel, p22 phox (green) was co-immunostained with caveolin-3 (red) to demonstrate sarcolemmal localization (yellow).

Techniques Used: Isolation, Staining


Structured Review

Santa Cruz Biotechnology rabbit anti p22 phox
Delayed inflammatory protein expression at 4 weeks and 4 months after contusive rat SCI. Protein expression of microglia/macrophages marker Iba-1 and NADPH oxidase component <t>p22</t> <t>PHOX</t> were analyzed using Western blotting at 4 weeks and 4 months post-injury. ( A-B ) Representative Western blots of Iba-1, p22 PHOX and the loading control at 4 weeks (A) and 4 months (B) after SCI. ( C ) Western blot analysis of Iba-1 and p22 PHOX protein expression indicated a significant increase at 4 weeks after SCI followed by a prolonged upregulation for up to 4 months post-injury. Bars represent mean ± SEM. * p < 0.05 compared with sham group. n = 4-6 rats per time point.
Rabbit Anti P22 Phox, supplied by Santa Cruz Biotechnology, 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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1) Product Images from "Delayed expression of cell cycle proteins contributes to astroglial scar formation and chronic inflammation after rat spinal cord contusion"

Article Title: Delayed expression of cell cycle proteins contributes to astroglial scar formation and chronic inflammation after rat spinal cord contusion

Journal: Journal of Neuroinflammation

doi: 10.1186/1742-2094-9-169

Delayed inflammatory protein expression at 4 weeks and 4 months after contusive rat SCI. Protein expression of microglia/macrophages marker Iba-1 and NADPH oxidase component p22 PHOX were analyzed using Western blotting at 4 weeks and 4 months post-injury. ( A-B ) Representative Western blots of Iba-1, p22 PHOX and the loading control at 4 weeks (A) and 4 months (B) after SCI. ( C ) Western blot analysis of Iba-1 and p22 PHOX protein expression indicated a significant increase at 4 weeks after SCI followed by a prolonged upregulation for up to 4 months post-injury. Bars represent mean ± SEM. * p < 0.05 compared with sham group. n = 4-6 rats per time point.
Figure Legend Snippet: Delayed inflammatory protein expression at 4 weeks and 4 months after contusive rat SCI. Protein expression of microglia/macrophages marker Iba-1 and NADPH oxidase component p22 PHOX were analyzed using Western blotting at 4 weeks and 4 months post-injury. ( A-B ) Representative Western blots of Iba-1, p22 PHOX and the loading control at 4 weeks (A) and 4 months (B) after SCI. ( C ) Western blot analysis of Iba-1 and p22 PHOX protein expression indicated a significant increase at 4 weeks after SCI followed by a prolonged upregulation for up to 4 months post-injury. Bars represent mean ± SEM. * p < 0.05 compared with sham group. n = 4-6 rats per time point.

Techniques Used: Expressing, Marker, Western Blot

The later upregulation of cell cycle proteins is associated with activated microglial/macrophages. The expression of PCNA, cyclin D1 and E was evaluated by immunohistochemistry at 1.5 mm caudal to the epicenter of sham or injured spinal cord at 1 month post-injury. ( A-D ) PCNA expression was undetectable in sham, but was strongly upregulated in OX42 + microglia/macrophages. ( E-H ) The membrane bound component of the NADPH oxidase enzyme, p22, co-localized with PCNA + microglia/macrophages. ( I-P ) Large numbers of cyclin E + and D1 + cells in the injured coronal sections were co-labeled with Iba-1 + or OX42 + microglia/macrophages. Scale bars = 500 μm.
Figure Legend Snippet: The later upregulation of cell cycle proteins is associated with activated microglial/macrophages. The expression of PCNA, cyclin D1 and E was evaluated by immunohistochemistry at 1.5 mm caudal to the epicenter of sham or injured spinal cord at 1 month post-injury. ( A-D ) PCNA expression was undetectable in sham, but was strongly upregulated in OX42 + microglia/macrophages. ( E-H ) The membrane bound component of the NADPH oxidase enzyme, p22, co-localized with PCNA + microglia/macrophages. ( I-P ) Large numbers of cyclin E + and D1 + cells in the injured coronal sections were co-labeled with Iba-1 + or OX42 + microglia/macrophages. Scale bars = 500 μm.

Techniques Used: Expressing, Immunohistochemistry, Labeling

Delayed systemic treatment with CR8 reduces chronic inflammatory protein expression at 4 months post-SCI. ( A ) Double-labeling immunohistochemistry revealed increased expression of PCNA (b) and OX42 (e), and their co-localization (h) in the injured tissue at 4 months post-injury. Delayed systemic CR8 treatment reduced immunoreactivity in both PCNA and OX42 (c, f, i). ( B ) SCI induced the increase of expression of Iba-1 and p22 PHOX in contrast to sham tissue (c-d), whereas these increases were attenuated by CR8 treatment (e-f). Scale bars = 500 μm.
Figure Legend Snippet: Delayed systemic treatment with CR8 reduces chronic inflammatory protein expression at 4 months post-SCI. ( A ) Double-labeling immunohistochemistry revealed increased expression of PCNA (b) and OX42 (e), and their co-localization (h) in the injured tissue at 4 months post-injury. Delayed systemic CR8 treatment reduced immunoreactivity in both PCNA and OX42 (c, f, i). ( B ) SCI induced the increase of expression of Iba-1 and p22 PHOX in contrast to sham tissue (c-d), whereas these increases were attenuated by CR8 treatment (e-f). Scale bars = 500 μm.

Techniques Used: Expressing, Labeling, Immunohistochemistry


Structured Review

Santa Cruz Biotechnology rabbit anti p22 phox

Rabbit Anti P22 Phox, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti p22 phox/product/Santa Cruz Biotechnology
Average 95 stars, based on 1 article reviews
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Images

1) Product Images from "Flaxseed Oil Containing α -Linolenic Acid Ester of Plant Sterol Improved Atherosclerosis in ApoE Deficient Mice"

Article Title: Flaxseed Oil Containing α -Linolenic Acid Ester of Plant Sterol Improved Atherosclerosis in ApoE Deficient Mice

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2015/958217


Figure Legend Snippet:

Techniques Used:

Effects of FO+ALA-PS on mRNA and protein expressions of p22 phox (a-b), p47 phox (c-d), p67 phox (e-f), and gp91 phox (g-h) in aorta of mice. Total RNA was extracted from aortas of mice by Trizol. p22 phox , p47 phox , p67 phox , and gp91 phox mRNA expressions were analyzed by real-time RT-PCR. The mRNA of β -actin was quantified as an endogenous control. Aortic lysates were prepared and immunoblotted with corresponding antibody, respectively. Blotting with anti- β -actin was used as a protein loading control. p22phox, p47phox, p67phox, and gp91phox are presented as fold change relative to control. Values are given as mean ± standard deviation of the mean ( n = 3). a P < 0.05 versus the control; b P < 0.05 versus the HFD group; c P < 0.05 versus the FO group.
Figure Legend Snippet: Effects of FO+ALA-PS on mRNA and protein expressions of p22 phox (a-b), p47 phox (c-d), p67 phox (e-f), and gp91 phox (g-h) in aorta of mice. Total RNA was extracted from aortas of mice by Trizol. p22 phox , p47 phox , p67 phox , and gp91 phox mRNA expressions were analyzed by real-time RT-PCR. The mRNA of β -actin was quantified as an endogenous control. Aortic lysates were prepared and immunoblotted with corresponding antibody, respectively. Blotting with anti- β -actin was used as a protein loading control. p22phox, p47phox, p67phox, and gp91phox are presented as fold change relative to control. Values are given as mean ± standard deviation of the mean ( n = 3). a P < 0.05 versus the control; b P < 0.05 versus the HFD group; c P < 0.05 versus the FO group.

Techniques Used: Quantitative RT-PCR, Standard Deviation

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  • 86
    Santa Cruz Biotechnology rabbit anti p22 phox
    a. Schematic of experiment for disrupting ETS2 in primary monocytes and differentiating monocyte-derived macrophages under chronic inflammatory conditions. b. Macrophage cytokine secretion following ETS2 disruption. Heatmap shows log2 fold-change of cytokine concentrations in the supernatants of ETS2 -edited macrophages relative to unedited macrophages transfected with a non-targeting control gRNA-containing RNP (NTC). n=9, Wilcoxon matched-pairs test, two-tailed. c. Histogram depicting phagocytosis of fluorescently-labelled zymosan particles by ETS2 -edited and unedited macrophages (left). Data representative of one of seven donors. Phagocytosis index in ETS2 -edited and unedited macrophages (calculated as product of proportion and mean fluorescence intensity of phagocytosing cells; right). Plot depicts log2 fold-change in phagocytosis index for ETS2 -edited macrophages relative to unedited cells (Wilcoxon signed-rank test, two-tailed; data represent mean±SEM). d. Production of ROS by ETS2 -edited and unedited inflammatory macrophages (measured in relative light units; left). Data representative of one of six donors. Western blot for gp91 <t>phox</t> , <t>p22</t> phox , and EROS expression in ETS2 -edited and unedited macrophages (right). Data representative of one of three donors. e. Differentially-expressed genes in ETS2 -edited versus unedited inflammatory macrophages (limma with voom transformation, n=8). f. Gene set enrichment analysis (fGSEA) of differentially-expressed genes between ETS2 -edited and unedited inflammatory macrophages. Results of selected Gene Ontology Biological Pathways shown. Dot size represents P -value and colour denotes normalised enrichment score (NES). g. Enrichment of differentially-expressed genes following deletion of the disease-associated chr21q22 locus (upregulated genes, top; downregulated genes, bottom) in ETS2 -edited versus unedited macrophages. * P < 0.05, ** P < 0.01.
    Rabbit Anti P22 Phox, supplied by Santa Cruz Biotechnology, 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/rabbit anti p22 phox/product/Santa Cruz Biotechnology
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    95
    Santa Cruz Biotechnology rabbit polyclonal antibodies against p22 phox
    Nox-4 (a) and <t>p22</t> <t>phox</t> (b) protein expressions in the liver. m/m : Misty; Veh: vehicle-treated db/db mice; K-100: Kangen-karyu 100 mg/kg body weight-treated db/db mice; K-200: Kangen-karyu 200 mg/kg body weight-treated db/db mice. Bars represent the means ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001 versus vehicle-treated db/db mouse values.
    Rabbit Polyclonal Antibodies Against P22 Phox, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal antibodies against p22 phox/product/Santa Cruz Biotechnology
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal antibodies against p22 phox - by Bioz Stars, 2024-03
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    86
    Santa Cruz Biotechnology rabbit p22 phox
    ( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to <t>p22</t> <t>phox</t> (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.
    Rabbit P22 Phox, supplied by Santa Cruz Biotechnology, 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/rabbit p22 phox/product/Santa Cruz Biotechnology
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    rabbit p22 phox - by Bioz Stars, 2024-03
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    95
    Santa Cruz Biotechnology rabbit polyclonal anti p22 phox
    ( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to <t>p22</t> <t>phox</t> (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.
    Rabbit Polyclonal Anti P22 Phox, supplied by Santa Cruz Biotechnology, 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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    86
    Santa Cruz Biotechnology rabbit antibodies against p22 phox
    A. Representative western blots of all NADPH oxidase subunits from TA muscles of 18–19 day old WT and mdx mice. B. Pooled results from densitometry for the subunits showing mdx values normalized to WT (represented by the blue dotted line). The proteins gp91 <t>phox</t> , p67 phox and rac1 were all significantly increased in mdx muscle compared to WT, whereas <t>p22</t> phox and p47 phox were not different (n = 4 samples for each group).
    Rabbit Antibodies Against P22 Phox, supplied by Santa Cruz Biotechnology, 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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    Image Search Results


    a. Schematic of experiment for disrupting ETS2 in primary monocytes and differentiating monocyte-derived macrophages under chronic inflammatory conditions. b. Macrophage cytokine secretion following ETS2 disruption. Heatmap shows log2 fold-change of cytokine concentrations in the supernatants of ETS2 -edited macrophages relative to unedited macrophages transfected with a non-targeting control gRNA-containing RNP (NTC). n=9, Wilcoxon matched-pairs test, two-tailed. c. Histogram depicting phagocytosis of fluorescently-labelled zymosan particles by ETS2 -edited and unedited macrophages (left). Data representative of one of seven donors. Phagocytosis index in ETS2 -edited and unedited macrophages (calculated as product of proportion and mean fluorescence intensity of phagocytosing cells; right). Plot depicts log2 fold-change in phagocytosis index for ETS2 -edited macrophages relative to unedited cells (Wilcoxon signed-rank test, two-tailed; data represent mean±SEM). d. Production of ROS by ETS2 -edited and unedited inflammatory macrophages (measured in relative light units; left). Data representative of one of six donors. Western blot for gp91 phox , p22 phox , and EROS expression in ETS2 -edited and unedited macrophages (right). Data representative of one of three donors. e. Differentially-expressed genes in ETS2 -edited versus unedited inflammatory macrophages (limma with voom transformation, n=8). f. Gene set enrichment analysis (fGSEA) of differentially-expressed genes between ETS2 -edited and unedited inflammatory macrophages. Results of selected Gene Ontology Biological Pathways shown. Dot size represents P -value and colour denotes normalised enrichment score (NES). g. Enrichment of differentially-expressed genes following deletion of the disease-associated chr21q22 locus (upregulated genes, top; downregulated genes, bottom) in ETS2 -edited versus unedited macrophages. * P < 0.05, ** P < 0.01.

    Journal: bioRxiv

    Article Title: A disease-associated gene desert orchestrates macrophage inflammatory responses via ETS2

    doi: 10.1101/2023.05.05.539522

    Figure Lengend Snippet: a. Schematic of experiment for disrupting ETS2 in primary monocytes and differentiating monocyte-derived macrophages under chronic inflammatory conditions. b. Macrophage cytokine secretion following ETS2 disruption. Heatmap shows log2 fold-change of cytokine concentrations in the supernatants of ETS2 -edited macrophages relative to unedited macrophages transfected with a non-targeting control gRNA-containing RNP (NTC). n=9, Wilcoxon matched-pairs test, two-tailed. c. Histogram depicting phagocytosis of fluorescently-labelled zymosan particles by ETS2 -edited and unedited macrophages (left). Data representative of one of seven donors. Phagocytosis index in ETS2 -edited and unedited macrophages (calculated as product of proportion and mean fluorescence intensity of phagocytosing cells; right). Plot depicts log2 fold-change in phagocytosis index for ETS2 -edited macrophages relative to unedited cells (Wilcoxon signed-rank test, two-tailed; data represent mean±SEM). d. Production of ROS by ETS2 -edited and unedited inflammatory macrophages (measured in relative light units; left). Data representative of one of six donors. Western blot for gp91 phox , p22 phox , and EROS expression in ETS2 -edited and unedited macrophages (right). Data representative of one of three donors. e. Differentially-expressed genes in ETS2 -edited versus unedited inflammatory macrophages (limma with voom transformation, n=8). f. Gene set enrichment analysis (fGSEA) of differentially-expressed genes between ETS2 -edited and unedited inflammatory macrophages. Results of selected Gene Ontology Biological Pathways shown. Dot size represents P -value and colour denotes normalised enrichment score (NES). g. Enrichment of differentially-expressed genes following deletion of the disease-associated chr21q22 locus (upregulated genes, top; downregulated genes, bottom) in ETS2 -edited versus unedited macrophages. * P < 0.05, ** P < 0.01.

    Article Snippet: Western blotting was performed as described previously using the following primary antibodies: rabbit anti-gp91 phox , rabbit anti-p22 phox (both Santa Cruz), rabbit anti-C17ORF62/EROS (Atlas), rabbit anti-actin (Abcam).

    Techniques: Derivative Assay, Transfection, Two Tailed Test, Fluorescence, Western Blot, Expressing, Transformation Assay

    Nox-4 (a) and p22 phox (b) protein expressions in the liver. m/m : Misty; Veh: vehicle-treated db/db mice; K-100: Kangen-karyu 100 mg/kg body weight-treated db/db mice; K-200: Kangen-karyu 200 mg/kg body weight-treated db/db mice. Bars represent the means ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001 versus vehicle-treated db/db mouse values.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Evaluation of Effects of Chinese Prescription Kangen-karyu on Diabetes-Induced Alterations such as Oxidative Stress and Apoptosis in the Liver of Type 2 Diabetic db/db Mice

    doi: 10.1155/2012/143489

    Figure Lengend Snippet: Nox-4 (a) and p22 phox (b) protein expressions in the liver. m/m : Misty; Veh: vehicle-treated db/db mice; K-100: Kangen-karyu 100 mg/kg body weight-treated db/db mice; K-200: Kangen-karyu 200 mg/kg body weight-treated db/db mice. Bars represent the means ± S.E.M. * P < 0.05, ** P < 0.01, *** P < 0.001 versus vehicle-treated db/db mouse values.

    Article Snippet: Rabbit polyclonal antibodies against p22 phox , NF- κ Bp65, nuclear factor erythroid 2-related factor 2 (Nrf-2), heme oxygenase-1 (HO-1), cytochrome c , Bax, and mouse monoclonal antibodies against cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), Bcl-2, and histone were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

    Techniques:

    ( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.

    Journal: PLoS ONE

    Article Title: Reactive Oxygen Species Regulate the Levels of Dual Oxidase (Duox1-2) in Human Neuroblastoma Cells

    doi: 10.1371/journal.pone.0034405

    Figure Lengend Snippet: ( A ) Identification of NOX expressed in neuroblastoma (SK-N-BE) and colon carcinoma (Caco-2) cells. Total RNA was extracted with Trizol, reverse transcribed and analyzed by PCR (see ) with specific primers to NOX1, NOX2 and NOX4 (left) or NOX3 and NOX5 (right). The number of cycles was 35. ( B ) Cells were transfected by electroporation with two different (45 and 46) siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 48h after transfection, total proteins were extracted and subjected to immunoblot analysis of p22 phox . The histograms show the mean +/- SEM values relative to scramble obtained by densitometric analysis of p22 phox bands normalized for α-tubulin of three independent experiments. *p< 0.01 vs scramble. ( C ) 24h after transfection cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. Total proteins were extracted and subjected to immunoblot analysis of DUOX. The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) obtained by densitometric analysis of DUOX bands normalized for α-tubulin of three independent experiments. * p< 0.01 vs Ctr. ( D, E ) 24h after transfection with a mix of the two p22 phox siRNA, cells were incubated in medium containing 0.2% FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min, mRNA was extracted and DUOX1 and DUOX2 mRNA levels were analyzed by RT-PCR as described in . The histograms show the mean +/- SEM values relative to samples not stimulated with PDGF (Ctr) of three independent experiments. * p<0.05 vs Ctr; ** p<0.05 vs PDGF stimulated scramble.

    Article Snippet: DUOX 1 and 2 proteins were detected with a rabbit polyclonal antibody raised against the peptide sequence ETELTPQRLQC located inside the first intracellular loop of human DUOX1; the specificity of the antibodies was tested using human primary thyroid cells as positive control and pre-immune serum as negative control (not shown); goat anti gp91 phox (NOX2) and rabbit p22 phox or SOD1 polyclonal antibodies were purchased by Santa Cruz Biotechnology (USA); The filters were also probed with an anti α-tubulin antibody (Sigma, USA).

    Techniques: Transfection, Electroporation, Western Blot, Incubation, Reverse Transcription Polymerase Chain Reaction

    ( A ) Cells were incubated 18h in medium containing 0.2% FBS, loaded with 10µM DCHF-DA in the presence or absence of the intracellular calcium chelator, BAPTA-AM (10µM), and then stimulated with 15ng/ml of PDGF as described in . ROS levels were measured fluorimetrically at the time intervals indicated. Ca ++ -independent ROS were measured in presence of BAPTA-AM. Ca ++ -dependent ROS levels were derived from the assays performed in the presence or absence of BAPTA-AM. Total levels of ROS induced by PDGF were also measured in the absence (Total) or presence of the NADPH oxidase inhibitor AEBSF. Values are Mean +/- SEM of three independent experiments performed in triplicate. * p< 0.01 and ** p< 0.05 vs not stimulated; § p<0.01 vs the corresponding time point of Total (Ctr) curve. ( B ) Cells were transfected by electroporation with siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 24h after transfection cells were incubated in medium containing 0.2%FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. An aliquot of cell medium was collected and analyzed for H 2 O 2 levels as described in . The histograms show the mean +/- SEM values of three independent experiments. * p< 0.01 vs Ctr. ** p< 0.01 vs PDGF stimulated scramble.

    Journal: PLoS ONE

    Article Title: Reactive Oxygen Species Regulate the Levels of Dual Oxidase (Duox1-2) in Human Neuroblastoma Cells

    doi: 10.1371/journal.pone.0034405

    Figure Lengend Snippet: ( A ) Cells were incubated 18h in medium containing 0.2% FBS, loaded with 10µM DCHF-DA in the presence or absence of the intracellular calcium chelator, BAPTA-AM (10µM), and then stimulated with 15ng/ml of PDGF as described in . ROS levels were measured fluorimetrically at the time intervals indicated. Ca ++ -independent ROS were measured in presence of BAPTA-AM. Ca ++ -dependent ROS levels were derived from the assays performed in the presence or absence of BAPTA-AM. Total levels of ROS induced by PDGF were also measured in the absence (Total) or presence of the NADPH oxidase inhibitor AEBSF. Values are Mean +/- SEM of three independent experiments performed in triplicate. * p< 0.01 and ** p< 0.05 vs not stimulated; § p<0.01 vs the corresponding time point of Total (Ctr) curve. ( B ) Cells were transfected by electroporation with siRNA to p22 phox (siRNA p22 phox ) or control, scrambled siRNA (scramble) as described in . 24h after transfection cells were incubated in medium containing 0.2%FBS for 18h and then stimulated with 15ng/ml of PDGF for 15min. An aliquot of cell medium was collected and analyzed for H 2 O 2 levels as described in . The histograms show the mean +/- SEM values of three independent experiments. * p< 0.01 vs Ctr. ** p< 0.01 vs PDGF stimulated scramble.

    Article Snippet: DUOX 1 and 2 proteins were detected with a rabbit polyclonal antibody raised against the peptide sequence ETELTPQRLQC located inside the first intracellular loop of human DUOX1; the specificity of the antibodies was tested using human primary thyroid cells as positive control and pre-immune serum as negative control (not shown); goat anti gp91 phox (NOX2) and rabbit p22 phox or SOD1 polyclonal antibodies were purchased by Santa Cruz Biotechnology (USA); The filters were also probed with an anti α-tubulin antibody (Sigma, USA).

    Techniques: Incubation, Derivative Assay, Transfection, Electroporation

    A. Representative western blots of all NADPH oxidase subunits from TA muscles of 18–19 day old WT and mdx mice. B. Pooled results from densitometry for the subunits showing mdx values normalized to WT (represented by the blue dotted line). The proteins gp91 phox , p67 phox and rac1 were all significantly increased in mdx muscle compared to WT, whereas p22 phox and p47 phox were not different (n = 4 samples for each group).

    Journal: PLoS ONE

    Article Title: Skeletal Muscle NADPH Oxidase Is Increased and Triggers Stretch-Induced Damage in the mdx Mouse

    doi: 10.1371/journal.pone.0015354

    Figure Lengend Snippet: A. Representative western blots of all NADPH oxidase subunits from TA muscles of 18–19 day old WT and mdx mice. B. Pooled results from densitometry for the subunits showing mdx values normalized to WT (represented by the blue dotted line). The proteins gp91 phox , p67 phox and rac1 were all significantly increased in mdx muscle compared to WT, whereas p22 phox and p47 phox were not different (n = 4 samples for each group).

    Article Snippet: Membranes were probed using mouse antibodies against rac1 (1∶500) (Cytoskeleton, CO), gp91 phox (1∶1000) and p67 phox (1∶500) (BD Biosciences, San Jose, CA), and rabbit antibodies against p22 phox (1∶500) and p47 phox (1∶500) (Santa Cruz, CA).

    Techniques: Western Blot

    Isolated mdx muscle fibers from the FDB muscle were immunostained using antibodies against the various NADPH oxidase subunits. Nuclei are stained by DAPI (blue). In the bottom right panel, p22 phox (green) was co-immunostained with caveolin-3 (red) to demonstrate sarcolemmal localization (yellow).

    Journal: PLoS ONE

    Article Title: Skeletal Muscle NADPH Oxidase Is Increased and Triggers Stretch-Induced Damage in the mdx Mouse

    doi: 10.1371/journal.pone.0015354

    Figure Lengend Snippet: Isolated mdx muscle fibers from the FDB muscle were immunostained using antibodies against the various NADPH oxidase subunits. Nuclei are stained by DAPI (blue). In the bottom right panel, p22 phox (green) was co-immunostained with caveolin-3 (red) to demonstrate sarcolemmal localization (yellow).

    Article Snippet: Membranes were probed using mouse antibodies against rac1 (1∶500) (Cytoskeleton, CO), gp91 phox (1∶1000) and p67 phox (1∶500) (BD Biosciences, San Jose, CA), and rabbit antibodies against p22 phox (1∶500) and p47 phox (1∶500) (Santa Cruz, CA).

    Techniques: Isolation, Staining