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Santa Cruz Biotechnology nqo1
BL153 upregulated renal PGC-1 α and <t>NQO1</t> expression. PGC-1 α and NQO1 expression was detected by western blot assay. n = 5; * P
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1) Product Images from "Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model"

Article Title: Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2013/367040

BL153 upregulated renal PGC-1 α and NQO1 expression. PGC-1 α and NQO1 expression was detected by western blot assay. n = 5; * P
Figure Legend Snippet: BL153 upregulated renal PGC-1 α and NQO1 expression. PGC-1 α and NQO1 expression was detected by western blot assay. n = 5; * P

Techniques Used: Pyrolysis Gas Chromatography, Expressing, Western Blot

2) Product Images from "Delayed treatment with oleanolic acid attenuates tubulointerstitial fibrosis in chronic cyclosporine nephropathy through Nrf2/HO-1 signaling"

Article Title: Delayed treatment with oleanolic acid attenuates tubulointerstitial fibrosis in chronic cyclosporine nephropathy through Nrf2/HO-1 signaling

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-12-50

Effects of oleanolic acid (OA) on nuclear/total Nrf2, Keap1, HO-1 and NQO1 expressions in chronic CsA nephropathy. (A) Representative Western blot showing the effects of OA on nuclear/total Nrf2, Keap1, HO-1 and NQO1 expression in chronic CsA nephropathy. (B) Quantitative analyses for total Nrf2/β-actin. There were no significant differences identified by quantitative analysis for immunoblotting of total Nrf2 among the experimental groups. (C) Quantitative analyses for nuclear/total Nrf2. There was increased nuclear/total Nrf2 in the CsA + OA compared with CsA. *p
Figure Legend Snippet: Effects of oleanolic acid (OA) on nuclear/total Nrf2, Keap1, HO-1 and NQO1 expressions in chronic CsA nephropathy. (A) Representative Western blot showing the effects of OA on nuclear/total Nrf2, Keap1, HO-1 and NQO1 expression in chronic CsA nephropathy. (B) Quantitative analyses for total Nrf2/β-actin. There were no significant differences identified by quantitative analysis for immunoblotting of total Nrf2 among the experimental groups. (C) Quantitative analyses for nuclear/total Nrf2. There was increased nuclear/total Nrf2 in the CsA + OA compared with CsA. *p

Techniques Used: Western Blot, Expressing

3) Product Images from "PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy"

Article Title: PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy

Journal: Chemistry & Biology

doi: 10.1016/j.chembiol.2014.09.019

PMI Stabilizes Nrf2 and Upregulates P62 Expression Activating Mitophagy (A) Structures of compounds 1 (sulforaphane) and 2 (PMI); CD is the concentration of compound causing a doubling of the control level of NQO1 enzymatic activity. (B) Western blot to show induction of Nrf2-dependent gene products versus time in Hepa1c1c7 cells (cytoplasmic) exposed to 10 μM PMI. (C) Induction of NQO1 (NAD(P)H dependent quinone oxidoreductase-1) by compounds 1 (○) and 2 (●). (D) Western blots to demonstrate Nrf2 stabilization in cells treated with either PMI (10 μM) or, sulforaphane (1 μM) versus time (E) RT-PCR analysis for estimation of p62 mRNA levels in MEFs following treatment with PMI versus time. Values are presented as arbitrary units normalized to 18 s RNA levels for each sample, n ≥ 3. (F) Western blot to demonstrate P62 expression in MEF cells treated with DMSO vehicle control, 10 μM PMI, or 1 μM sulforaphane for 24 hr. Β-actin is shown as a loading control. (G) Graph shows P62:β-actin ratio band density analysis, n = 3. (H) Representative confocal images of β-subunit staining to highlight mitochondrial density in MEF cells treated with DMSO vehicle control or 10 μM PMI for 24 hr. (I) Graph shows average mitochondrial area as a percentage of cell size, n ≥ 50. (J) Western blot to demonstrate reduction in MTCO1 levels following 4 hr FCCP or 24 hr PMI exposure. (K) Graph shows MTCO1:Tubulin ratio band density analysis normalized to control, n = 3. All values are mean ± SEM, ∗ p
Figure Legend Snippet: PMI Stabilizes Nrf2 and Upregulates P62 Expression Activating Mitophagy (A) Structures of compounds 1 (sulforaphane) and 2 (PMI); CD is the concentration of compound causing a doubling of the control level of NQO1 enzymatic activity. (B) Western blot to show induction of Nrf2-dependent gene products versus time in Hepa1c1c7 cells (cytoplasmic) exposed to 10 μM PMI. (C) Induction of NQO1 (NAD(P)H dependent quinone oxidoreductase-1) by compounds 1 (○) and 2 (●). (D) Western blots to demonstrate Nrf2 stabilization in cells treated with either PMI (10 μM) or, sulforaphane (1 μM) versus time (E) RT-PCR analysis for estimation of p62 mRNA levels in MEFs following treatment with PMI versus time. Values are presented as arbitrary units normalized to 18 s RNA levels for each sample, n ≥ 3. (F) Western blot to demonstrate P62 expression in MEF cells treated with DMSO vehicle control, 10 μM PMI, or 1 μM sulforaphane for 24 hr. Β-actin is shown as a loading control. (G) Graph shows P62:β-actin ratio band density analysis, n = 3. (H) Representative confocal images of β-subunit staining to highlight mitochondrial density in MEF cells treated with DMSO vehicle control or 10 μM PMI for 24 hr. (I) Graph shows average mitochondrial area as a percentage of cell size, n ≥ 50. (J) Western blot to demonstrate reduction in MTCO1 levels following 4 hr FCCP or 24 hr PMI exposure. (K) Graph shows MTCO1:Tubulin ratio band density analysis normalized to control, n = 3. All values are mean ± SEM, ∗ p

Techniques Used: Expressing, Concentration Assay, Activity Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining

4) Product Images from "CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes"

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206152

Effects of rimonabant on Nrf2 and its downstream gene expression and AMPK phosphorylation in rat livers. (A) Hepatic Nrf2 and antioxidant-responsive element (ARE)-mediated NQO1, HO-1, GSTA, and TRNRD1 gene expression were determined by RT-PCR, normalized for all samples to ribosomal RNA (18S) level, and expressed as fold change compared to LETO control rats (LETO Con). Representative western blots for NQO1, HO-1, and β-actin (B) and p-AMPK, AMPK, and β-actin (D). The density of signal was quantified and normalized by β-actin (C) or AMPK (E). Data are expressed as mean ± SEM (n = 4–5 per group). *P
Figure Legend Snippet: Effects of rimonabant on Nrf2 and its downstream gene expression and AMPK phosphorylation in rat livers. (A) Hepatic Nrf2 and antioxidant-responsive element (ARE)-mediated NQO1, HO-1, GSTA, and TRNRD1 gene expression were determined by RT-PCR, normalized for all samples to ribosomal RNA (18S) level, and expressed as fold change compared to LETO control rats (LETO Con). Representative western blots for NQO1, HO-1, and β-actin (B) and p-AMPK, AMPK, and β-actin (D). The density of signal was quantified and normalized by β-actin (C) or AMPK (E). Data are expressed as mean ± SEM (n = 4–5 per group). *P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

5) Product Images from "Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2"

Article Title: Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2

Journal: Carcinogenesis

doi: 10.1093/carcin/bgn095

The Nrf2-dependent defense response is upregulated in lung tumors and cancer cell lines. cDNAs from human lung tissues of normal and various stages of cancers were subjected to real-time reverse transcription–PCR for detection of NQO1 mRNA expression.
Figure Legend Snippet: The Nrf2-dependent defense response is upregulated in lung tumors and cancer cell lines. cDNAs from human lung tissues of normal and various stages of cancers were subjected to real-time reverse transcription–PCR for detection of NQO1 mRNA expression.

Techniques Used: Polymerase Chain Reaction, Expressing

6) Product Images from "An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil"

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil

Journal: Nutrition & Metabolism

doi: 10.1186/s12986-017-0225-z

Effect on protein expression of Nrf2 targets. Relative Protein expression of GPX ( a ), HO-1 ( b ) and NQO1 ( c ) in the liver of rats fed diets with either soybean oil or salmon oil with various vitamin E concentrations. Bars represent relative protein level expressed as fold of control (diet with soybean oil and 25 mg/kg vitamin E) and are means ± SD from 12 animals per group. Representative immunoblots specific to GPX ( a ), HO-1 ( b ), NQO1 ( c ) and ß-actin as respective loading control are shown for one animal per group
Figure Legend Snippet: Effect on protein expression of Nrf2 targets. Relative Protein expression of GPX ( a ), HO-1 ( b ) and NQO1 ( c ) in the liver of rats fed diets with either soybean oil or salmon oil with various vitamin E concentrations. Bars represent relative protein level expressed as fold of control (diet with soybean oil and 25 mg/kg vitamin E) and are means ± SD from 12 animals per group. Representative immunoblots specific to GPX ( a ), HO-1 ( b ), NQO1 ( c ) and ß-actin as respective loading control are shown for one animal per group

Techniques Used: Expressing, Western Blot

7) Product Images from "Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling"

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling

Journal: American Journal of Translational Research

doi:

Cellular expressions of Nrf2 + cells and NQO1 + cells in kidney parenchyma by day 7 after IR procedure. A. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in glomeruli among the three groups. B. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in medulla among the three groups. C. Analytic results of number of Nrf2 + cells in glomeruli, * vs. other groups with different symbols (†, ‡), p
Figure Legend Snippet: Cellular expressions of Nrf2 + cells and NQO1 + cells in kidney parenchyma by day 7 after IR procedure. A. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in glomeruli among the three groups. B. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in medulla among the three groups. C. Analytic results of number of Nrf2 + cells in glomeruli, * vs. other groups with different symbols (†, ‡), p

Techniques Used:

8) Product Images from "Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet"

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet

Journal: Nutrients

doi: 10.3390/nu8120799

Myricetin activated the hepatic Nrf2 pathway and normalized expressions of genes involved in oxidative stress. ( A ) Protein expression of hepatic nuclear Nrf2; ( B ) protein expression of hepatic cytosolic Nrf2; ( C ) protein expression of hepatic NQO1 and HO-1; and ( D ) relative expression of genes involved in oxidative stress. Values were presented as mean ± SEM ( n = 8). Con, control diet; CM, control diet with additional 0.12% myricetin; HFD, high-fat diet; HM, high-fat diet with additional 0.12% myricetin. * p
Figure Legend Snippet: Myricetin activated the hepatic Nrf2 pathway and normalized expressions of genes involved in oxidative stress. ( A ) Protein expression of hepatic nuclear Nrf2; ( B ) protein expression of hepatic cytosolic Nrf2; ( C ) protein expression of hepatic NQO1 and HO-1; and ( D ) relative expression of genes involved in oxidative stress. Values were presented as mean ± SEM ( n = 8). Con, control diet; CM, control diet with additional 0.12% myricetin; HFD, high-fat diet; HM, high-fat diet with additional 0.12% myricetin. * p

Techniques Used: Expressing

9) Product Images from "Sulforaphane Inhibits Mitochondrial Permeability Transition and Oxidative Stress"

Article Title: Sulforaphane Inhibits Mitochondrial Permeability Transition and Oxidative Stress

Journal: Free radical biology & medicine

doi: 10.1016/j.freeradbiomed.2011.09.017

NAD(P)H quinone oxidoreductase 1 (NQO1) immunoreactivity in liver homogenates from vehicle- and sulforaphane (SFP) treated rats. A. Representative immunoblots for NQO1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). B. Densitometric ratios for NQO1/VDAC
Figure Legend Snippet: NAD(P)H quinone oxidoreductase 1 (NQO1) immunoreactivity in liver homogenates from vehicle- and sulforaphane (SFP) treated rats. A. Representative immunoblots for NQO1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). B. Densitometric ratios for NQO1/VDAC

Techniques Used: Western Blot

10) Product Images from "Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas"

Article Title: Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas

Journal: Cancer cell

doi: 10.1016/j.ccell.2017.10.011

p62 Ablation Attenuates Pancreatitis-Accelerated Neoplastic Progression (A) Quantification of ADM and PanIN1 density, amylase and Ki67 staining of pancreatic sections of indicated 5-week-old mice (n = 5). (B) H E staining, amylase IHC, amylase and CK19 co-IF, and Sox9 IHC of pancreatic sections from indicated 5-week-old mice. Scale bars: 25 µm. (C) Kaplan-Meier survival curves of indicated mouse strains (n = 10). (D) Q-RT-PCR analysis of mRNA in acinar and ductal cell fractions from indicated 5-week-old mice (n = 5). (E) NQO1, MDM2 and HES1 IHC of pancreatic sections from indicated 5-week-old mice. Scale bars: 25 µm. (F) p53 IB analysis of pancreatic lysates from 5-week-old mice of indicated genotypes. (G) Frequency of ALDH expression in EpCAM + cells from 8-week-old Kras G12D (n = 3), Kras G12D ; Ikkα Δpan (n = 7), and Kras G12D ; Ikkα/p62 Δpan (n = 4) mice. (H) Sphere-forming capacity of isolated ALDH + .
Figure Legend Snippet: p62 Ablation Attenuates Pancreatitis-Accelerated Neoplastic Progression (A) Quantification of ADM and PanIN1 density, amylase and Ki67 staining of pancreatic sections of indicated 5-week-old mice (n = 5). (B) H E staining, amylase IHC, amylase and CK19 co-IF, and Sox9 IHC of pancreatic sections from indicated 5-week-old mice. Scale bars: 25 µm. (C) Kaplan-Meier survival curves of indicated mouse strains (n = 10). (D) Q-RT-PCR analysis of mRNA in acinar and ductal cell fractions from indicated 5-week-old mice (n = 5). (E) NQO1, MDM2 and HES1 IHC of pancreatic sections from indicated 5-week-old mice. Scale bars: 25 µm. (F) p53 IB analysis of pancreatic lysates from 5-week-old mice of indicated genotypes. (G) Frequency of ALDH expression in EpCAM + cells from 8-week-old Kras G12D (n = 3), Kras G12D ; Ikkα Δpan (n = 7), and Kras G12D ; Ikkα/p62 Δpan (n = 4) mice. (H) Sphere-forming capacity of isolated ALDH + .

Techniques Used: Staining, Mouse Assay, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation

11) Product Images from "CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes"

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes

Journal: PLoS ONE

doi: 10.1371/journal.pone.0206152

Effects of rimonabant on Nrf2 and its downstream gene expression and AMPK phosphorylation in rat livers. (A) Hepatic Nrf2 and antioxidant-responsive element (ARE)-mediated NQO1, HO-1, GSTA, and TRNRD1 gene expression were determined by RT-PCR, normalized for all samples to ribosomal RNA (18S) level, and expressed as fold change compared to LETO control rats (LETO Con). Representative western blots for NQO1, HO-1, and β-actin (B) and p-AMPK, AMPK, and β-actin (D). The density of signal was quantified and normalized by β-actin (C) or AMPK (E). Data are expressed as mean ± SEM (n = 4–5 per group). *P
Figure Legend Snippet: Effects of rimonabant on Nrf2 and its downstream gene expression and AMPK phosphorylation in rat livers. (A) Hepatic Nrf2 and antioxidant-responsive element (ARE)-mediated NQO1, HO-1, GSTA, and TRNRD1 gene expression were determined by RT-PCR, normalized for all samples to ribosomal RNA (18S) level, and expressed as fold change compared to LETO control rats (LETO Con). Representative western blots for NQO1, HO-1, and β-actin (B) and p-AMPK, AMPK, and β-actin (D). The density of signal was quantified and normalized by β-actin (C) or AMPK (E). Data are expressed as mean ± SEM (n = 4–5 per group). *P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

12) Product Images from "Royal jelly attenuates cadmium-induced nephrotoxicity in male mice"

Article Title: Royal jelly attenuates cadmium-induced nephrotoxicity in male mice

Journal: Scientific Reports

doi: 10.1038/s41598-019-42368-7

Western blot (WB) analysis and densitometric quantification of NF-κB and Nrf2 and its putative target proteins HO-1 and NQO1 in the kidney of mice intoxicated with CdCl 2 and pretreated with royal jelly (RJ). Data (mean ± SD of triplicate assays). β-actin was used as the loading control. Molecular weight of proteins is indicated at the right-hand side. Full-length blots/gels are presented in Supplementary Fig. S1 . a p
Figure Legend Snippet: Western blot (WB) analysis and densitometric quantification of NF-κB and Nrf2 and its putative target proteins HO-1 and NQO1 in the kidney of mice intoxicated with CdCl 2 and pretreated with royal jelly (RJ). Data (mean ± SD of triplicate assays). β-actin was used as the loading control. Molecular weight of proteins is indicated at the right-hand side. Full-length blots/gels are presented in Supplementary Fig. S1 . a p

Techniques Used: Western Blot, Mouse Assay, Molecular Weight

13) Product Images from "Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways"

Article Title: Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways

Journal: PLoS ONE

doi: 10.1371/journal.pone.0185374

Effects of TQ on NQO1 expression. NQO1 mRNA expression in liver (A) and soleus muscle (B). (C) Western blot images of NQO1 and β-actin protein in liver (D) Western blot images of NQO1 protein in soleus muscle. β-tubulin was used as a loading control. Statistical analysis (A and B): one-way ANOVA followed by Sidak post-test (p≤0.05). qPCR results are means ± SEM (n = 8–12 mice per treatment group). Western blot images are representative of combined liver and soleus muscle lysates from n = 10–12 mice per treatment group. LFD: low fat diet, HFD: high fat diet, TQ: thymoquinone.
Figure Legend Snippet: Effects of TQ on NQO1 expression. NQO1 mRNA expression in liver (A) and soleus muscle (B). (C) Western blot images of NQO1 and β-actin protein in liver (D) Western blot images of NQO1 protein in soleus muscle. β-tubulin was used as a loading control. Statistical analysis (A and B): one-way ANOVA followed by Sidak post-test (p≤0.05). qPCR results are means ± SEM (n = 8–12 mice per treatment group). Western blot images are representative of combined liver and soleus muscle lysates from n = 10–12 mice per treatment group. LFD: low fat diet, HFD: high fat diet, TQ: thymoquinone.

Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Mouse Assay

14) Product Images from "NQO1-mediated tumor-selective lethality and radiosensitization for head and neck cancer"

Article Title: NQO1-mediated tumor-selective lethality and radiosensitization for head and neck cancer

Journal: Molecular cancer therapeutics

doi: 10.1158/1535-7163.MCT-15-0765

Expression of NQO1 and Catalase in HNC cell lines
Figure Legend Snippet: Expression of NQO1 and Catalase in HNC cell lines

Techniques Used: Expressing

Functional implications of NQO1 expression in response to β-lap
Figure Legend Snippet: Functional implications of NQO1 expression in response to β-lap

Techniques Used: Functional Assay, Expressing

Functional consequences of reduced NQO1 expression and mechanisms of β-lap-induced NQO1-dependent cell death
Figure Legend Snippet: Functional consequences of reduced NQO1 expression and mechanisms of β-lap-induced NQO1-dependent cell death

Techniques Used: Functional Assay, Expressing

Expression of NQO1 and Catalase in HNC and adjacent normal tissues
Figure Legend Snippet: Expression of NQO1 and Catalase in HNC and adjacent normal tissues

Techniques Used: Expressing

15) Product Images from "Poly(ADP-ribose) polymerase-1 Modulates Nrf2-dependent Transcription"

Article Title: Poly(ADP-ribose) polymerase-1 Modulates Nrf2-dependent Transcription

Journal: Free radical biology & medicine

doi: 10.1016/j.freeradbiomed.2013.10.806

PARP-1 augments Nrf2 binding to the ARE and upregulates expression of Nrf2 target genes. (A) PARP-1 enhances the binding of endogenous Nrf2 to the NQO1-ARE. Biotinylated NQO1-ARE DNA was incubated with whole cell lysates from MDA-MB-231 cells expressing
Figure Legend Snippet: PARP-1 augments Nrf2 binding to the ARE and upregulates expression of Nrf2 target genes. (A) PARP-1 enhances the binding of endogenous Nrf2 to the NQO1-ARE. Biotinylated NQO1-ARE DNA was incubated with whole cell lysates from MDA-MB-231 cells expressing

Techniques Used: Binding Assay, Expressing, Incubation, Multiple Displacement Amplification

PARP-1 upregualtes the transcriptional activity of Nrf2. (A) PARP-1 promotes the NQO1-ARE-dependent luciferase activity. MDA-MB-231 cells were co-transfected with expression vectors for either wild-type or mutant NQO1-ARE-dependent firefly luciferase
Figure Legend Snippet: PARP-1 upregualtes the transcriptional activity of Nrf2. (A) PARP-1 promotes the NQO1-ARE-dependent luciferase activity. MDA-MB-231 cells were co-transfected with expression vectors for either wild-type or mutant NQO1-ARE-dependent firefly luciferase

Techniques Used: Activity Assay, Luciferase, Multiple Displacement Amplification, Transfection, Expressing, Mutagenesis

PARP-1 binds the NQO1-ARE. (A) Sequences of wild-type (ARE) and mutant NQO1-ARE (mARE) used in this study. Nucleotides labeled in red are mutated in mARE. (B) Silver stain of proteins pulled down by the biotinylated ARE or mARE pull-downs/streptavidin
Figure Legend Snippet: PARP-1 binds the NQO1-ARE. (A) Sequences of wild-type (ARE) and mutant NQO1-ARE (mARE) used in this study. Nucleotides labeled in red are mutated in mARE. (B) Silver stain of proteins pulled down by the biotinylated ARE or mARE pull-downs/streptavidin

Techniques Used: Mutagenesis, Labeling, Silver Staining

16) Product Images from "Leflunomide Attenuates Oxidative Stress in Fetal Human Lung Endothelial Cells via Superoxide Dismutase 2 and Catalase"

Article Title: Leflunomide Attenuates Oxidative Stress in Fetal Human Lung Endothelial Cells via Superoxide Dismutase 2 and Catalase

Journal: Biochemical and biophysical research communications

doi: 10.1016/j.bbrc.2018.07.149

Leflunomide-treated HPAECs display increased catalase, NQO1, and SOD2 expression HPAECs were treated with leflunomide (L) at doses of 5 (L5) or 10 (L10) μM or DMSO for 24 h, and the cells were harvested for real-time RT-PCR and immunoblot analyses. A–E. Real-time RT-PCR analyses-based determination of catalase (A), HO-1 (B), NQO1 (C), SOD1 (D), and SOD2 (E) mRNA-expression levels. F. Determination of catalase, HO-1, SOD1, SOD2, and NQO1 protein levels by immunoblotting. G-K. Quantification and normalization of catalase (G), HO-1 (H), NQO1 (I), SOD1 (J), and SOD2 (K) band intensities to those of β-actin. The data shown are representative of three independent experiments. Values are presented as the mean ± SD (n = 3/group). One-way ANOVA showed an effect of L5 and L10 for catalase, NQO1, and SOD2 mRNA and protein expression. Significant differences between DMSO-, L5- and L10-treated cells are indicated by *, p
Figure Legend Snippet: Leflunomide-treated HPAECs display increased catalase, NQO1, and SOD2 expression HPAECs were treated with leflunomide (L) at doses of 5 (L5) or 10 (L10) μM or DMSO for 24 h, and the cells were harvested for real-time RT-PCR and immunoblot analyses. A–E. Real-time RT-PCR analyses-based determination of catalase (A), HO-1 (B), NQO1 (C), SOD1 (D), and SOD2 (E) mRNA-expression levels. F. Determination of catalase, HO-1, SOD1, SOD2, and NQO1 protein levels by immunoblotting. G-K. Quantification and normalization of catalase (G), HO-1 (H), NQO1 (I), SOD1 (J), and SOD2 (K) band intensities to those of β-actin. The data shown are representative of three independent experiments. Values are presented as the mean ± SD (n = 3/group). One-way ANOVA showed an effect of L5 and L10 for catalase, NQO1, and SOD2 mRNA and protein expression. Significant differences between DMSO-, L5- and L10-treated cells are indicated by *, p

Techniques Used: Expressing, Quantitative RT-PCR

Leflunomide treated HPAECs display decreased H 2 O 2 levels and increased NQO1 and SOD2 protein expression upon exposure to hyperoxia HPAECs were treated with leflunomide (L) at doses of 5 (L5) or 10 (L10) μM or DMSO and exposed to normoxia or hyperoxia for 24 h, and the cells were harvested for H 2 O 2 estimation (A), and real-time RT-PCR and immunoblot analyses. B–F. Real-time RT-PCR analyses-based determination of catalase (B), HO-1 (C), NQO1 (D), SOD1 (E), and SOD2 (F) mRNA-expression levels. G. Determination of catalase, HO-1, NQO1, SOD1 and SOD2 protein levels by immunoblotting. H-L. Quantification and normalization of catalase (H), HO-1 (I), NQO1 (J), SOD1 (K), and SOD2 (L) band intensities to those of β-actin. The data shown are representative of three independent experiments. Values are presented as the mean ± SD (n = 3/group). Two-way ANOVA showed an effect of leflunomide and hyperoxia on the dependent variables, H 2 O 2 level, and HO-1 and NQO1 expression, and an interaction between leflunomide and hyperoxia for the dependent variables, H 2 O 2 level, and NQO1 and SOD2 expression. Significant differences between vehicle and leflunomide treated cells exposed to normoxia are indicated by p, p
Figure Legend Snippet: Leflunomide treated HPAECs display decreased H 2 O 2 levels and increased NQO1 and SOD2 protein expression upon exposure to hyperoxia HPAECs were treated with leflunomide (L) at doses of 5 (L5) or 10 (L10) μM or DMSO and exposed to normoxia or hyperoxia for 24 h, and the cells were harvested for H 2 O 2 estimation (A), and real-time RT-PCR and immunoblot analyses. B–F. Real-time RT-PCR analyses-based determination of catalase (B), HO-1 (C), NQO1 (D), SOD1 (E), and SOD2 (F) mRNA-expression levels. G. Determination of catalase, HO-1, NQO1, SOD1 and SOD2 protein levels by immunoblotting. H-L. Quantification and normalization of catalase (H), HO-1 (I), NQO1 (J), SOD1 (K), and SOD2 (L) band intensities to those of β-actin. The data shown are representative of three independent experiments. Values are presented as the mean ± SD (n = 3/group). Two-way ANOVA showed an effect of leflunomide and hyperoxia on the dependent variables, H 2 O 2 level, and HO-1 and NQO1 expression, and an interaction between leflunomide and hyperoxia for the dependent variables, H 2 O 2 level, and NQO1 and SOD2 expression. Significant differences between vehicle and leflunomide treated cells exposed to normoxia are indicated by p, p

Techniques Used: Expressing, Quantitative RT-PCR

17) Product Images from "Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling"

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling

Journal: American Journal of Translational Research

doi:

Cellular expressions of Nrf2 + cells and NQO1 + cells in kidney parenchyma by day 7 after IR procedure. A. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in glomeruli among the three groups. B. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in medulla among the three groups. C. Analytic results of number of Nrf2 + cells in glomeruli, * vs. other groups with different symbols (†, ‡), p
Figure Legend Snippet: Cellular expressions of Nrf2 + cells and NQO1 + cells in kidney parenchyma by day 7 after IR procedure. A. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in glomeruli among the three groups. B. Illustrating the IF microscopic finding (200x) for identification of Nrf2 + cells (green color) and NQO1 + cells (red color) in medulla among the three groups. C. Analytic results of number of Nrf2 + cells in glomeruli, * vs. other groups with different symbols (†, ‡), p

Techniques Used:

18) Product Images from "Kalopanacis Cortex extract-capped gold nanoparticles activate NRF2 signaling and ameliorate damage in human neuronal SH-SY5Y cells exposed to oxygen–glucose deprivation and reoxygenation"

Article Title: Kalopanacis Cortex extract-capped gold nanoparticles activate NRF2 signaling and ameliorate damage in human neuronal SH-SY5Y cells exposed to oxygen–glucose deprivation and reoxygenation

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S138178

Scheme illustrating the neuroprotective properties of KC-GNs against OGD-induced injury in human neuronal cells. Abbreviations: ARE, antioxidant response element; HO-1, heme oxygenase-1; KC-GN, Kalopanacis Cortex extract-capped gold nanoparticle; NQO1, NAD(P)H quinone dehydrogenase 1; NRF2, nuclear factor erythroid 2-related factor 2; OGD, oxygen–glucose deprivation/reoxygenation; ROS, reactive oxygen species.
Figure Legend Snippet: Scheme illustrating the neuroprotective properties of KC-GNs against OGD-induced injury in human neuronal cells. Abbreviations: ARE, antioxidant response element; HO-1, heme oxygenase-1; KC-GN, Kalopanacis Cortex extract-capped gold nanoparticle; NQO1, NAD(P)H quinone dehydrogenase 1; NRF2, nuclear factor erythroid 2-related factor 2; OGD, oxygen–glucose deprivation/reoxygenation; ROS, reactive oxygen species.

Techniques Used:

KC-GNs activate NRF2/ARE signaling and induce the expression of HO-1 and NQO1. Notes: Cells were pretreated with KC-GNs or ci-GNs for 1 h before OGD/R treatment. ( A ) Fixed cells were stained with PI and an anti-NRF2 antibody, followed by incubation with an FITC-conjugated anti-rabbit IgG secondary antibody. The samples were observed by confocal microscopy. Scale bar =20 μm. ( B ) Western blot was used to detect the NRF2 protein, with TBP used as the internal control. ( C ) Cells transfected with the ARE–luciferase reporter plasmid were incubated with KC-GNs for 1 h and then exposed to OGD/R. Equal amounts of the cell extract were assayed for dual-luciferase activity. ( D ) Expression of the HO-1 and NQO1 proteins was examined by Western blot. ( E ) Cells were transfected with the HO-1 promoter reporter plasmid. Each bar represents the mean ± standard error of three independent experiments per group. ** P
Figure Legend Snippet: KC-GNs activate NRF2/ARE signaling and induce the expression of HO-1 and NQO1. Notes: Cells were pretreated with KC-GNs or ci-GNs for 1 h before OGD/R treatment. ( A ) Fixed cells were stained with PI and an anti-NRF2 antibody, followed by incubation with an FITC-conjugated anti-rabbit IgG secondary antibody. The samples were observed by confocal microscopy. Scale bar =20 μm. ( B ) Western blot was used to detect the NRF2 protein, with TBP used as the internal control. ( C ) Cells transfected with the ARE–luciferase reporter plasmid were incubated with KC-GNs for 1 h and then exposed to OGD/R. Equal amounts of the cell extract were assayed for dual-luciferase activity. ( D ) Expression of the HO-1 and NQO1 proteins was examined by Western blot. ( E ) Cells were transfected with the HO-1 promoter reporter plasmid. Each bar represents the mean ± standard error of three independent experiments per group. ** P

Techniques Used: Expressing, Staining, Incubation, Confocal Microscopy, Western Blot, Transfection, Luciferase, Plasmid Preparation, Activity Assay

19) Product Images from "The Effect of Aging on Acetaminophen Pharmacokinetics, Toxicity and Nrf2 in Fischer 344 Rats"

Article Title: The Effect of Aging on Acetaminophen Pharmacokinetics, Toxicity and Nrf2 in Fischer 344 Rats

Journal: The Journals of Gerontology Series A: Biological Sciences and Medical Sciences

doi: 10.1093/gerona/glt095

Protein expression of Cytochrome P450 2E1 (CYP2E1), NAD(P)H quinone oxireductase 1 (NQO1) and glutathione synthesis regulating proteins of young (6 mo) and old (26 mo) rats 4 h after saline (solid bar) or acetaminophen (APAP; open bars) intraperitoneal injection. (A) CYP2E1, (B) NQO1, (C) Glutathione cysteine ligase–Catalytic subunit (GCLC), (D) Glutathione cysteine ligase–Modulatory subunit (GCLM), and (E) activated Nuclear factor (erythroid-derived 2)-like 2 (Nrf2). All data are presented as mean arbitrary densitometric values relative to young control as 1 ± SEM of n = 5–9 animals per group with immunoblot respresentative of target protein and loading control results below graph. Statistically significant differences comparing against corresponding young group: * p
Figure Legend Snippet: Protein expression of Cytochrome P450 2E1 (CYP2E1), NAD(P)H quinone oxireductase 1 (NQO1) and glutathione synthesis regulating proteins of young (6 mo) and old (26 mo) rats 4 h after saline (solid bar) or acetaminophen (APAP; open bars) intraperitoneal injection. (A) CYP2E1, (B) NQO1, (C) Glutathione cysteine ligase–Catalytic subunit (GCLC), (D) Glutathione cysteine ligase–Modulatory subunit (GCLM), and (E) activated Nuclear factor (erythroid-derived 2)-like 2 (Nrf2). All data are presented as mean arbitrary densitometric values relative to young control as 1 ± SEM of n = 5–9 animals per group with immunoblot respresentative of target protein and loading control results below graph. Statistically significant differences comparing against corresponding young group: * p

Techniques Used: Expressing, Injection, Derivative Assay

20) Product Images from "Basal autophagy maintains pancreatic acinar cell homeostasis and protein synthesis and prevents ER stress"

Article Title: Basal autophagy maintains pancreatic acinar cell homeostasis and protein synthesis and prevents ER stress

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1519384112

The Atg7 Δpan phenotype is only partially rescued by additional p62 ablation. ( A ) Amylase (Amy2a) ( Left ), Nqo1 and Gstm1 ( Right ) mRNA amounts in pancreata of indicated genotypes. Results are means ± SEM n = 3 mice per condition. * P
Figure Legend Snippet: The Atg7 Δpan phenotype is only partially rescued by additional p62 ablation. ( A ) Amylase (Amy2a) ( Left ), Nqo1 and Gstm1 ( Right ) mRNA amounts in pancreata of indicated genotypes. Results are means ± SEM n = 3 mice per condition. * P

Techniques Used: Mouse Assay

21) Product Images from "Comparative Study of 17-AAG and NVP-AUY922 in Pancreatic and Colorectal Cancer Cells: Are There Common Determinants of Sensitivity?"

Article Title: Comparative Study of 17-AAG and NVP-AUY922 in Pancreatic and Colorectal Cancer Cells: Are There Common Determinants of Sensitivity?

Journal: Translational Oncology

doi: 10.1016/j.tranon.2014.08.001

Pharmacological NQO1 inhibition. Cell proliferation assays. (A) Pancreatic and colorectal carcinoma cells were treated with ES936 for 30 minutes before treatment with DMSO (control), 0.1 μM or 0.5 μM 17-AAG for 72 hours, and cell proliferation
Figure Legend Snippet: Pharmacological NQO1 inhibition. Cell proliferation assays. (A) Pancreatic and colorectal carcinoma cells were treated with ES936 for 30 minutes before treatment with DMSO (control), 0.1 μM or 0.5 μM 17-AAG for 72 hours, and cell proliferation

Techniques Used: Inhibition

Comparison of NQO1 protein levels and activity. (A) Western blot analyses were performed with total cellular extracts to determine basal NQO1, Hsp70, or β-actin levels in the cell lines or primary cultures indicated. NQO1 activity assays. (B)
Figure Legend Snippet: Comparison of NQO1 protein levels and activity. (A) Western blot analyses were performed with total cellular extracts to determine basal NQO1, Hsp70, or β-actin levels in the cell lines or primary cultures indicated. NQO1 activity assays. (B)

Techniques Used: Activity Assay, Western Blot

Biologic NQO1 inhibition. IMIM-PC-2 cells were nontransfected (control) or transfected with a control siRNA (scrambled sequence) or a specific siRNA against NQO1, before treatment with DMSO or 0.5 μM 17-AAG for 72 hours. (A) EGFR, Hsp70, NQO1,
Figure Legend Snippet: Biologic NQO1 inhibition. IMIM-PC-2 cells were nontransfected (control) or transfected with a control siRNA (scrambled sequence) or a specific siRNA against NQO1, before treatment with DMSO or 0.5 μM 17-AAG for 72 hours. (A) EGFR, Hsp70, NQO1,

Techniques Used: Inhibition, Transfection, Sequencing

22) Product Images from "Human Cytomegalovirus Induces Multiple Means To Combat Reactive Oxygen Species ▿"

Article Title: Human Cytomegalovirus Induces Multiple Means To Combat Reactive Oxygen Species ▿

Journal: Journal of Virology

doi: 10.1128/JVI.05572-11

HCMV infection increases RNA levels of specific Nrf2 target genes. Real-time PCR analysis of GCLC, HO-1, and NQO1 RNA levels in HCMV-infected cells allowed comparisons to mock-infected cells. Total mRNA was extracted from HFs at 0, 24, 48, 60, 72, 96,
Figure Legend Snippet: HCMV infection increases RNA levels of specific Nrf2 target genes. Real-time PCR analysis of GCLC, HO-1, and NQO1 RNA levels in HCMV-infected cells allowed comparisons to mock-infected cells. Total mRNA was extracted from HFs at 0, 24, 48, 60, 72, 96,

Techniques Used: Infection, Real-time Polymerase Chain Reaction

23) Product Images from "Natural antioxidants exhibit chemopreventive characteristics through the regulation of CNC-bZip transcription factors in estrogen-induced breast carcinogenesis"

Article Title: Natural antioxidants exhibit chemopreventive characteristics through the regulation of CNC-bZip transcription factors in estrogen-induced breast carcinogenesis

Journal: Journal of biochemical and molecular toxicology

doi: 10.1002/jbt.21594

Antioxidants reverse E2-mediated decrease in Phase-II detoxifying enzymes, NQO1 and SOD3, at both mRNA and protein levels. (A) NQO1 mRNA expression levels in MCF-10A cells treated with 50 nM E2, 50 μM Res, Res + E2, 1 mM VC or VC + E2 for 24 hr;
Figure Legend Snippet: Antioxidants reverse E2-mediated decrease in Phase-II detoxifying enzymes, NQO1 and SOD3, at both mRNA and protein levels. (A) NQO1 mRNA expression levels in MCF-10A cells treated with 50 nM E2, 50 μM Res, Res + E2, 1 mM VC or VC + E2 for 24 hr;

Techniques Used: Expressing

Nrf3 negatively regulates Nrf2 and Nrf2-dependent gene NQO1 and Nrf3 interacts with Nrf2. (A) MCF-10A cells were transfected with 20 nmol/l of scrambled siRNA or siNrf3 for 48 hr, and Western blot analysis was carried out using Nrf3 antibody. The same
Figure Legend Snippet: Nrf3 negatively regulates Nrf2 and Nrf2-dependent gene NQO1 and Nrf3 interacts with Nrf2. (A) MCF-10A cells were transfected with 20 nmol/l of scrambled siRNA or siNrf3 for 48 hr, and Western blot analysis was carried out using Nrf3 antibody. The same

Techniques Used: Transfection, Western Blot

24) Product Images from "Piper betle induces phase I II genes through Nrf2/ARE signaling pathway in mouse embryonic fibroblasts derived from wild type and Nrf2 knockout cells"

Article Title: Piper betle induces phase I II genes through Nrf2/ARE signaling pathway in mouse embryonic fibroblasts derived from wild type and Nrf2 knockout cells

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/1472-6882-14-72

Western blot of NQO1, HO-1 and SOD1 proteins in WT and N0 cells treated with PB (5 and 10 μg/ml). The blots shown are examples of three separate experiments (A) from 20 μg cytosolic proteins and relative intensities over β-actin for NQO1 (B) , HO-1 (C) and SOD1 (D) protein expression after 24 h treatment of PB. SFN treatment at 5 μM was used as positive control. Data represent ± S.E.M from three independent experiments. a P
Figure Legend Snippet: Western blot of NQO1, HO-1 and SOD1 proteins in WT and N0 cells treated with PB (5 and 10 μg/ml). The blots shown are examples of three separate experiments (A) from 20 μg cytosolic proteins and relative intensities over β-actin for NQO1 (B) , HO-1 (C) and SOD1 (D) protein expression after 24 h treatment of PB. SFN treatment at 5 μM was used as positive control. Data represent ± S.E.M from three independent experiments. a P

Techniques Used: Western Blot, Expressing, Positive Control

ARE-driven luciferase activities after treatment with PB in MEFs WT and N0 cells. The cells were transfected with the luciferase reporter plasmid containing the NQO1 ARE and were treated with PB (5 and 10 μg/ml) or SFN (5 μM) for 24 h. Measurement of luciferase activities was performed using the Dual luciferase activities and Firefly luciferase levels were normalized to Renilla luciferase levels. Data represent ± S.E.M from four independent experiments. a P
Figure Legend Snippet: ARE-driven luciferase activities after treatment with PB in MEFs WT and N0 cells. The cells were transfected with the luciferase reporter plasmid containing the NQO1 ARE and were treated with PB (5 and 10 μg/ml) or SFN (5 μM) for 24 h. Measurement of luciferase activities was performed using the Dual luciferase activities and Firefly luciferase levels were normalized to Renilla luciferase levels. Data represent ± S.E.M from four independent experiments. a P

Techniques Used: Luciferase, Transfection, Plasmid Preparation

Transcript levels for Nrf2, NQO1 and SOD1 genes following 5 and 10 μg/ml of PB treatment for 24-h. RNAs were isolated and reverse transcribed to cDNA, then amplified by real-time PCR detection system to measure mRNA levels for beta actin, Nrf2 (A) , NQO1 (B) and SOD1 (C) , HO-1 (D) and GSTA1 (E) . Target genes were normalized to beta actin. Data represent ± S.E.M from three independent experiments. a P
Figure Legend Snippet: Transcript levels for Nrf2, NQO1 and SOD1 genes following 5 and 10 μg/ml of PB treatment for 24-h. RNAs were isolated and reverse transcribed to cDNA, then amplified by real-time PCR detection system to measure mRNA levels for beta actin, Nrf2 (A) , NQO1 (B) and SOD1 (C) , HO-1 (D) and GSTA1 (E) . Target genes were normalized to beta actin. Data represent ± S.E.M from three independent experiments. a P

Techniques Used: Isolation, Amplification, Real-time Polymerase Chain Reaction

25) Product Images from "Brg1-mediated Nrf2/HO-1 pathway activation alleviates hepatic ischemia–reperfusion injury"

Article Title: Brg1-mediated Nrf2/HO-1 pathway activation alleviates hepatic ischemia–reperfusion injury

Journal: Cell Death & Disease

doi: 10.1038/cddis.2017.236

The role of Brg1 in AML12 cells subjected to H/R injury. ( a ) Western blot analysis showed the cellular Brg1 and HO-1 protein expression in AML12 hepatocytes subjected to hypoxia for 12 h and reoxygenation for 0 (H12R0), 2 (H12R2), 4 (H12R4), 6 (H12R6), 8 (H12R8), 12 (H12R12), 16 (H12R16) and 24 h (H12R24) before sample collection in comparison with the cells cultured for the same time as control (namely, C12, C14, C16, C20, C24, C28 and C36, representing 12–36 h of culture). The proteins from the H/R groups and from the control groups were loaded in the same gel when performing western blotting assay and displayed in parallel to facilitate comparison. Representative images from one of three independent experiments were shown. ( b and c ) Quantitative measurement of band intensity in a by densitometry analysis. ( d ) Fluorescence immunostaining of DCF in cells with Brg1 overexpression using Brg1-Adv transfection, and elevated in cells with Brg1 knockdown using Brg1-siRNA transfection during H/R (H12R4) injury. Representative images from one of three independent experiments were shown. ( e ) Bar graph showing the change in DCF fluorescent intensity. ( f ) ELISA assay showed that 8-isoprostane level was decreased after Brg1-Adv treatment and increased by Brg1-siRNA transfection during H/R (H12R4) injury. ( g and h ) Western blot analysis showed the change of Brg1 and Nrf2 protein expression in AML12 cells, respectively, under condition of Brg1 overexpression or knockdown. Representative images were shown and quantitative measurements were performed. ( i and j ) Western blot and RT-PCR analysis showed the protein and mRNA level of HO-1 and NQO1 in response to Brg1 overexpression or knockdown during cell H/R (H12R4) injury. ( k ) HO-1 promoter-driven luciferase activity assay was performed and tBHQ (20 μ M) was used as Nrf2 nuclear translocation positive control. Data are mean±S.E.M. of three independent experiments each performed in triplicate. * P
Figure Legend Snippet: The role of Brg1 in AML12 cells subjected to H/R injury. ( a ) Western blot analysis showed the cellular Brg1 and HO-1 protein expression in AML12 hepatocytes subjected to hypoxia for 12 h and reoxygenation for 0 (H12R0), 2 (H12R2), 4 (H12R4), 6 (H12R6), 8 (H12R8), 12 (H12R12), 16 (H12R16) and 24 h (H12R24) before sample collection in comparison with the cells cultured for the same time as control (namely, C12, C14, C16, C20, C24, C28 and C36, representing 12–36 h of culture). The proteins from the H/R groups and from the control groups were loaded in the same gel when performing western blotting assay and displayed in parallel to facilitate comparison. Representative images from one of three independent experiments were shown. ( b and c ) Quantitative measurement of band intensity in a by densitometry analysis. ( d ) Fluorescence immunostaining of DCF in cells with Brg1 overexpression using Brg1-Adv transfection, and elevated in cells with Brg1 knockdown using Brg1-siRNA transfection during H/R (H12R4) injury. Representative images from one of three independent experiments were shown. ( e ) Bar graph showing the change in DCF fluorescent intensity. ( f ) ELISA assay showed that 8-isoprostane level was decreased after Brg1-Adv treatment and increased by Brg1-siRNA transfection during H/R (H12R4) injury. ( g and h ) Western blot analysis showed the change of Brg1 and Nrf2 protein expression in AML12 cells, respectively, under condition of Brg1 overexpression or knockdown. Representative images were shown and quantitative measurements were performed. ( i and j ) Western blot and RT-PCR analysis showed the protein and mRNA level of HO-1 and NQO1 in response to Brg1 overexpression or knockdown during cell H/R (H12R4) injury. ( k ) HO-1 promoter-driven luciferase activity assay was performed and tBHQ (20 μ M) was used as Nrf2 nuclear translocation positive control. Data are mean±S.E.M. of three independent experiments each performed in triplicate. * P

Techniques Used: Western Blot, Expressing, Cell Culture, Fluorescence, Immunostaining, Over Expression, Transfection, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Luciferase, Activity Assay, Translocation Assay, Positive Control

Expressions of Brg1, Nrf2 and Nrf2 downstream genes in the liver after hepatic I/R. ( a , b , c and d ) Western blot analysis showed that Brg1, nuclear Nrf2, HO-1 and NQO1 protein expressions were elevated in response to HIR in the liver at indicated time points. Representative images from one of three independent experiments were shown. Quantitative analyses of the results were also performed. ( e , f , g and h ) Transcript levels of Brg1, Nrf2, HO-1 and NQO1 in the liver in sham and HIR group were measured by RT-PCR. Each bar represents the mean±S.E.M. ( n=6 per group). * P
Figure Legend Snippet: Expressions of Brg1, Nrf2 and Nrf2 downstream genes in the liver after hepatic I/R. ( a , b , c and d ) Western blot analysis showed that Brg1, nuclear Nrf2, HO-1 and NQO1 protein expressions were elevated in response to HIR in the liver at indicated time points. Representative images from one of three independent experiments were shown. Quantitative analyses of the results were also performed. ( e , f , g and h ) Transcript levels of Brg1, Nrf2, HO-1 and NQO1 in the liver in sham and HIR group were measured by RT-PCR. Each bar represents the mean±S.E.M. ( n=6 per group). * P

Techniques Used: Western Blot, Reverse Transcription Polymerase Chain Reaction

Overexpression of Brg1 attenuated HIR injury via enhancing antioxidant enzyme. Animals were subjected to 70% liver warm ischemia for 60 min and live tissues were collected at 6 h after reperfusion. ( a ) Western blot analysis showed that Brg1 expression was increased in Brg1 overexpression (CMV-Brg1) mice compared to WT mice both in the sham and HIR groups. ( b ) Suzike’s injury score showed lower scores in CMV-Brg1 mice than in WT mice after HIR injury. ( c ) Representative H E staining images of liver collected from WT and CMV-Brg1 mice in the sham and HIR groups are shown. ( d and e ) Serum AST and ALT concentration showed an improved liver function in CMV-Brg1 mice after HIR injury. ( f ) ELISA analysis showed elevation of 8-isoprostane level was attenuated in CMV-Brg1 mice in response to HIR injury relative to that in the control. ( g ) ROS production measured by fluorescence intensity of DCF was reduced in CMV-Brg1 mice after HIR injury. ( h ) Immumohistochemical staining showed that liver HO-1 protein expression was elevated in response to HIR in CMV-Brg1 mice. ( i ) Quantitative analyses of the results from h were also performed. ( j ) Western blot analysis showed that liver NQO1 protein expression did not significantly change in CMV-Brg1 mice compared to WT mice. Each bar represents the mean±S.E.M. ( n=6 per group). * P
Figure Legend Snippet: Overexpression of Brg1 attenuated HIR injury via enhancing antioxidant enzyme. Animals were subjected to 70% liver warm ischemia for 60 min and live tissues were collected at 6 h after reperfusion. ( a ) Western blot analysis showed that Brg1 expression was increased in Brg1 overexpression (CMV-Brg1) mice compared to WT mice both in the sham and HIR groups. ( b ) Suzike’s injury score showed lower scores in CMV-Brg1 mice than in WT mice after HIR injury. ( c ) Representative H E staining images of liver collected from WT and CMV-Brg1 mice in the sham and HIR groups are shown. ( d and e ) Serum AST and ALT concentration showed an improved liver function in CMV-Brg1 mice after HIR injury. ( f ) ELISA analysis showed elevation of 8-isoprostane level was attenuated in CMV-Brg1 mice in response to HIR injury relative to that in the control. ( g ) ROS production measured by fluorescence intensity of DCF was reduced in CMV-Brg1 mice after HIR injury. ( h ) Immumohistochemical staining showed that liver HO-1 protein expression was elevated in response to HIR in CMV-Brg1 mice. ( i ) Quantitative analyses of the results from h were also performed. ( j ) Western blot analysis showed that liver NQO1 protein expression did not significantly change in CMV-Brg1 mice compared to WT mice. Each bar represents the mean±S.E.M. ( n=6 per group). * P

Techniques Used: Over Expression, Western Blot, Expressing, Mouse Assay, Staining, AST Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Fluorescence

26) Product Images from "IDH1 R132H mutation regulates glioma chemosensitivity through Nrf2 pathway"

Article Title: IDH1 R132H mutation regulates glioma chemosensitivity through Nrf2 pathway

Journal: Oncotarget

doi: 10.18632/oncotarget.15868

P53 was involved in the resistance mechanism of temozolomide mediated by Nrf2 and NQO1 P53 was significantly decreased in MT groups (A) . Levels of NQO1 and P53 were decreased when Nrf2 expression was reduced by siRNA-Nrf2 (1) in NC and WT over-expression cells (B) . After treated by 400 μM TMZ for 72 h, p53 was activated in NC, WT and MT groups, and p53 levels in MT groups were still lower than those in other groups (C) . *P
Figure Legend Snippet: P53 was involved in the resistance mechanism of temozolomide mediated by Nrf2 and NQO1 P53 was significantly decreased in MT groups (A) . Levels of NQO1 and P53 were decreased when Nrf2 expression was reduced by siRNA-Nrf2 (1) in NC and WT over-expression cells (B) . After treated by 400 μM TMZ for 72 h, p53 was activated in NC, WT and MT groups, and p53 levels in MT groups were still lower than those in other groups (C) . *P

Techniques Used: Expressing, Over Expression

Nrf2, NQO1 and MRP1 proteins level in IDH1 R132H overexpressing U87 cells and U251 cells Base level of NQO1 and MRP1 in U87 and U251 cells (A) . After 400 μM TMZ treatment for 72 h, the expression level of NQO1 and Nrf2 in U87 (B) and U251 cells was measured (C) . Western blotting experiments were performed in triplicate and representative images are shown. Quantification of protein expression is shown at the side or below the images. ***P
Figure Legend Snippet: Nrf2, NQO1 and MRP1 proteins level in IDH1 R132H overexpressing U87 cells and U251 cells Base level of NQO1 and MRP1 in U87 and U251 cells (A) . After 400 μM TMZ treatment for 72 h, the expression level of NQO1 and Nrf2 in U87 (B) and U251 cells was measured (C) . Western blotting experiments were performed in triplicate and representative images are shown. Quantification of protein expression is shown at the side or below the images. ***P

Techniques Used: Expressing, Western Blot

27) Product Images from "High glucose induces renal tubular epithelial injury via Sirt1/NF-kappaB/microR-29/Keap1 signal pathway"

Article Title: High glucose induces renal tubular epithelial injury via Sirt1/NF-kappaB/microR-29/Keap1 signal pathway

Journal: Journal of Translational Medicine

doi: 10.1186/s12967-015-0710-y

Effect of high glucose on renal tubule epithelia cell of HK-2 in vitro. Cells were triggered with doses of glucose (5.5, 15, 30 and 45) for 48 h. a Sirt1 activity was assessed. b NF-κB transcription activity was evaluated using luciferase reporter gene assay. c miR-29 expression was determined. d Western blot was performed to assess Keap1, GST, NQO1 and nuclear Nrf-2 expression. e Cell viability was evaluated using MTT assay. Data were presented as mean ± S.D. **P
Figure Legend Snippet: Effect of high glucose on renal tubule epithelia cell of HK-2 in vitro. Cells were triggered with doses of glucose (5.5, 15, 30 and 45) for 48 h. a Sirt1 activity was assessed. b NF-κB transcription activity was evaluated using luciferase reporter gene assay. c miR-29 expression was determined. d Western blot was performed to assess Keap1, GST, NQO1 and nuclear Nrf-2 expression. e Cell viability was evaluated using MTT assay. Data were presented as mean ± S.D. **P

Techniques Used: In Vitro, Activity Assay, Luciferase, Reporter Gene Assay, Expressing, Western Blot, MTT Assay

28) Product Images from "Nrf2-Dependent Induction of NQO1 in Mouse Aortic Endothelial Cells Overexpressing Catalase"

Article Title: Nrf2-Dependent Induction of NQO1 in Mouse Aortic Endothelial Cells Overexpressing Catalase

Journal: Free radical biology & medicine

doi: 10.1016/j.freeradbiomed.2011.04.020

Proposed molecular mechanism for the role of catalase overexpression on BaP-induced NQO1 expression.
Figure Legend Snippet: Proposed molecular mechanism for the role of catalase overexpression on BaP-induced NQO1 expression.

Techniques Used: Over Expression, Expressing

The effect of N-acetylcysteine on peroxides, and AhR and NQO1 expression. After pre-incubation with or without 10 nM N-acetylecysteine (NAC) for 1 hr, wild-type MAECs were treated with 1 μM BaP or culture medium alone (control) for 6 h (Western blot analysis) or for 1 h (peroxide measurement). The AhR and NQO1 protein levels were determined by Western blot analysis and quantitated (B, C respectively) relative to β-actin. Cellular peroxyl radicals were measured in terms of fluorescence intensity (FI) per μg protein using CDC-H2F diacetate as a probe. Values represent the mean ± SEM of five independent experiments. * P
Figure Legend Snippet: The effect of N-acetylcysteine on peroxides, and AhR and NQO1 expression. After pre-incubation with or without 10 nM N-acetylecysteine (NAC) for 1 hr, wild-type MAECs were treated with 1 μM BaP or culture medium alone (control) for 6 h (Western blot analysis) or for 1 h (peroxide measurement). The AhR and NQO1 protein levels were determined by Western blot analysis and quantitated (B, C respectively) relative to β-actin. Cellular peroxyl radicals were measured in terms of fluorescence intensity (FI) per μg protein using CDC-H2F diacetate as a probe. Values represent the mean ± SEM of five independent experiments. * P

Techniques Used: Expressing, Incubation, Western Blot, Fluorescence

The effect of H 2 O 2 on AhR and NQO1 expression, and Sp1 binding to the AhR promoter. Wild-type and hCat Tg MAECs were treated with or without 100 μM H 2 O 2 for 2 h, and then treated with or without 1 μM BaP for 4 h. (A) The amount of Sp1 bound to the AhR promoter was determined using ChIP analysis, and expressed as the ratio of input controls. AhR (B) and NQO1 (C) mRNA levels were determined by quantitative real-time RT-PCR and expressed relative to GAPDH mRNA. (D) The level of NQO1 protein was determined in whole cell extracts by Western blot analysis and quantitated relative to β-actin. Values represent the mean ± SEM of three independent experiments. * P
Figure Legend Snippet: The effect of H 2 O 2 on AhR and NQO1 expression, and Sp1 binding to the AhR promoter. Wild-type and hCat Tg MAECs were treated with or without 100 μM H 2 O 2 for 2 h, and then treated with or without 1 μM BaP for 4 h. (A) The amount of Sp1 bound to the AhR promoter was determined using ChIP analysis, and expressed as the ratio of input controls. AhR (B) and NQO1 (C) mRNA levels were determined by quantitative real-time RT-PCR and expressed relative to GAPDH mRNA. (D) The level of NQO1 protein was determined in whole cell extracts by Western blot analysis and quantitated relative to β-actin. Values represent the mean ± SEM of three independent experiments. * P

Techniques Used: Expressing, Binding Assay, Chromatin Immunoprecipitation, Quantitative RT-PCR, Western Blot

Knockdown of AhR reduces BaP-induced Nrf2 and NQO1 expression. Wild-type and hCat Tg MAECs were transfected with AhR siRNA or control siRNA, and then treated with or without 1 μM BaP as described in Materials and Methods. The level of AhR protein (A) was determined by Western blot analysis and expressed relative to β-actin (B). The mRNA level of AhR was determined by quantitative real-time RT-PCR and expressed relative to GAPDH mRNA (C). The mRNA levels of Nrf2 (D) and NQO1 (E) were determined by quantitative real-time RT-PCR; BaP-induced changes were expressed as a % of those observed in cells without BaP treatment. Values represent the mean ± SEM of five independent experiments. * P
Figure Legend Snippet: Knockdown of AhR reduces BaP-induced Nrf2 and NQO1 expression. Wild-type and hCat Tg MAECs were transfected with AhR siRNA or control siRNA, and then treated with or without 1 μM BaP as described in Materials and Methods. The level of AhR protein (A) was determined by Western blot analysis and expressed relative to β-actin (B). The mRNA level of AhR was determined by quantitative real-time RT-PCR and expressed relative to GAPDH mRNA (C). The mRNA levels of Nrf2 (D) and NQO1 (E) were determined by quantitative real-time RT-PCR; BaP-induced changes were expressed as a % of those observed in cells without BaP treatment. Values represent the mean ± SEM of five independent experiments. * P

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

Overexpression of catalase elevates BaP-induced NQO1 expression. Wild-type and hCat Tg MAECs at the 3 rd or 8 th -9 th passages were treated with or without 1 μM BaP in the presence or absence of 10 μM aminotriazole (AT) as indicated. The level of NQO1 protein was determined in whole cell extracts by Western blot analysis (A) and quantitated relative to β-actin (B). The level of NQO1 mRNA was determined by quantitative real-time RT-PCR and normalized to GAPDH mRNA (C). Values represent the mean ± SEM of five independent experiments. * P
Figure Legend Snippet: Overexpression of catalase elevates BaP-induced NQO1 expression. Wild-type and hCat Tg MAECs at the 3 rd or 8 th -9 th passages were treated with or without 1 μM BaP in the presence or absence of 10 μM aminotriazole (AT) as indicated. The level of NQO1 protein was determined in whole cell extracts by Western blot analysis (A) and quantitated relative to β-actin (B). The level of NQO1 mRNA was determined by quantitative real-time RT-PCR and normalized to GAPDH mRNA (C). Values represent the mean ± SEM of five independent experiments. * P

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

Overexpression of catalase increases BaP-induced Nrf2 accumulation in the nucleus and binding to the NQO1 promoter. Wild-type and hCat Tg MAECs were treated with or without 1 μM BaP. The level of nuclear Nrf2 was determined by Western blot analysis (A) and expressed relative to β-actin (B). Nrf2 binding to the NQO1 promoter was assessed by ChIP analysis (C). Genomic DNA bound to Nrf2 was recovered and quantified by quantitative real-time PCR using primer pairs specific for the NQO1-ARE region. Data are expressed relative to the quantity of input. Values represent the mean ± SEM of five independent experiments. * P
Figure Legend Snippet: Overexpression of catalase increases BaP-induced Nrf2 accumulation in the nucleus and binding to the NQO1 promoter. Wild-type and hCat Tg MAECs were treated with or without 1 μM BaP. The level of nuclear Nrf2 was determined by Western blot analysis (A) and expressed relative to β-actin (B). Nrf2 binding to the NQO1 promoter was assessed by ChIP analysis (C). Genomic DNA bound to Nrf2 was recovered and quantified by quantitative real-time PCR using primer pairs specific for the NQO1-ARE region. Data are expressed relative to the quantity of input. Values represent the mean ± SEM of five independent experiments. * P

Techniques Used: Over Expression, Binding Assay, Western Blot, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

Functional analysis of the NQO1 promoter. NQO1 promoter-luciferase reporter plasmids were constructed and cotransfected into wild-type and hCat Tg MAECs with a β-galactosidase expression plasmid as described in Materials and Methods. (A) Schematic diagram depicting wild-type (P1), ARE-mutated (P2), and XRE-mutated (P3) NQO1 promoter-luciferase constructs. Luciferase activity was measured using a luminescence assay and expressed relative to the luminosity of β-galactosidase assay (B). Values represent the mean ± SEM of five independent experiments. * P
Figure Legend Snippet: Functional analysis of the NQO1 promoter. NQO1 promoter-luciferase reporter plasmids were constructed and cotransfected into wild-type and hCat Tg MAECs with a β-galactosidase expression plasmid as described in Materials and Methods. (A) Schematic diagram depicting wild-type (P1), ARE-mutated (P2), and XRE-mutated (P3) NQO1 promoter-luciferase constructs. Luciferase activity was measured using a luminescence assay and expressed relative to the luminosity of β-galactosidase assay (B). Values represent the mean ± SEM of five independent experiments. * P

Techniques Used: Functional Assay, Luciferase, Construct, Expressing, Plasmid Preparation, Activity Assay, Luminescence Assay

Knockdown of Nrf2 reduces NQO1 expression. Wild-type and hCat Tg MAECs were transfected with Nrf2 siRNA or control siRNA, and then treated with or without 1 μM BaP as described in Materials and Methods. The level of Nrf2 (A) and NQO1 (C) proteins were determined by Western blot analysis and expressed relative to β-actin. The mRNA levels of Nrf2 (B) and NQO1 (D) were determined by quantitative real-time RT-PCR and expressed relative to GAPDH mRNA. Values represent the mean ± SEM of five independent experiments. * P
Figure Legend Snippet: Knockdown of Nrf2 reduces NQO1 expression. Wild-type and hCat Tg MAECs were transfected with Nrf2 siRNA or control siRNA, and then treated with or without 1 μM BaP as described in Materials and Methods. The level of Nrf2 (A) and NQO1 (C) proteins were determined by Western blot analysis and expressed relative to β-actin. The mRNA levels of Nrf2 (B) and NQO1 (D) were determined by quantitative real-time RT-PCR and expressed relative to GAPDH mRNA. Values represent the mean ± SEM of five independent experiments. * P

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

29) Product Images from "Reduced formation of depurinating estrogen–DNA adducts by sulforaphane or KEAP1 disruption in human mammary epithelial MCF-10A cells"

Article Title: Reduced formation of depurinating estrogen–DNA adducts by sulforaphane or KEAP1 disruption in human mammary epithelial MCF-10A cells

Journal: Carcinogenesis

doi: 10.1093/carcin/bgt246

Pathway for formation of estrogen depurinating DNA adducts. E 2 or E 1 can be oxidized to E 1/2 -3,4-quinone, which can bind to DNA to form 4-OHE 1/2 -1-N3Adenine or 4-OHE 1/2 -1-N7Guanine adducts. NQO1 reduces E 1/2 -3,4-quinones back to catechols and GST catalyzes
Figure Legend Snippet: Pathway for formation of estrogen depurinating DNA adducts. E 2 or E 1 can be oxidized to E 1/2 -3,4-quinone, which can bind to DNA to form 4-OHE 1/2 -1-N3Adenine or 4-OHE 1/2 -1-N7Guanine adducts. NQO1 reduces E 1/2 -3,4-quinones back to catechols and GST catalyzes

Techniques Used:

Effects of SFN on transcript, protein and activities of enzymes metabolizing E 2 or E 1 . ( A ) Effect of SFN on transcripts levels of estrogen metabolism enzymes. ( B ) Effect of SFN on protein levels of estrogen metabolism enzymes. ( C ) Effect of SFN on NQO1
Figure Legend Snippet: Effects of SFN on transcript, protein and activities of enzymes metabolizing E 2 or E 1 . ( A ) Effect of SFN on transcripts levels of estrogen metabolism enzymes. ( B ) Effect of SFN on protein levels of estrogen metabolism enzymes. ( C ) Effect of SFN on NQO1

Techniques Used:

Related Articles

Pyrolysis Gas Chromatography:

Article Title: Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model
Article Snippet: .. The latter were blocked with 5% milk, followed by incubation with the following antibodies: TNF-α , PGC-1α (Abcam, Cambridge, MA), PAI-1 (BD Bioscience, San Jose, CA), 3-NT (Millipore, Billerica, MA), 4-HNE (Alpha Diagnostic International, San Antonio, TX), HK II, β -actin, and NQO1 (SantaCruz Biotechnology, Santa Cruz, CA). .. After those membranes were washed with Tris-buffered saline (pH 7.2) containing 0.05% Tween 20 and incubated with the appropriate secondary antibodies.

Blocking Assay:

Article Title: Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways
Article Snippet: .. Following the transfer, membranes were blocked with TBST (10 mmol/l Tris-HCl pH 7.4, 150 mmol/l NaCl, and 0.1% Tween 20) containing 5% nonfat dry milk (blocking buffer) and incubated with the primary antibodies (diluted in blocking buffer overnight at 4°C) against SIRT-1 (Cell Signaling, cat. #9475), p-SIRT-1 (Cell Signaling, cat. #2314), Akt (Cell Signaling, cat. #9272), p-Akt (Cell Signaling, cat. #9271), AMPKα (Cell Signaling, cat. #5831), p-AMPKα (Cell Signaling, cat. #2535), NQO1 (Santa Cruz, cat. #sc-16464), β-actin (Cell Signaling, cat. #4970), and β-tubulin (Cell Signaling, cat. #2146). .. Membranes were incubated with goat anti-rabbit immunoglobulin (IgG) secondary antibody (Santa Cruz, cat. #sc-2030) for 1 h at room temperature, and washed 5 times.

Article Title: Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas
Article Snippet: Blots were incubated with 5% (w/v) nonfat dry milk in PBS with 0.05% (w/v) Tween 20 (Millipore Sigma, P9416) (PBST) at room temp for 1 hr to block nonspecific binding, and overnight at 4°C with primary antibodies in 3% BSA (w/v) in PBST and finally with HRP-conjugated secondary antibody in blocking buffer. .. The following antibodies were used: PCNA (BD Pharmingen, #555566), cyclin D1 (Santa Cruz, sc-8396), pERK (Cell Signaling, #9101), ERK (Cell Signaling, #9102), p62 (Progen, GP62-C), LC3B (Cell Signaling, #2775), Keap1 (Cell Signaling D6B12), NRF2 (Santa Cruz, sc-722), NQO1 (Santa Cruz, 271116), tubulin (Sigma, T9026), Hes1 (Santa Cruz, sc-25392), CK19 (Santa Cruz, sc-33111), p27 (Santa Cruz, sc-528), p53 (Cell Signaling, 2524), Sox9 (Santa Cruz, sc-20095), Ki67 (GeneTex, GTX 16667), MDM2 (Santa Cruz, sc-965), ALDH (Abcam, ab24343), p-Histone H2A.X (Ser 139) (Santa Cruz, sc-517348).

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: .. After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific). .. Immunoblot imaging was captured by using a BioSpectrum AC Imaging System (Ultra-Violet Products).

Electrophoresis:

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: Equal amounts of protein were separated by electrophoresis on 12% or 14% sodium dodecyl sulfate polyacrylamide gels and transferred to polyvinylidene difluoride membrane (Millipore, Marlborough, MA, USA). .. Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology).

Article Title: PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy
Article Snippet: Proteins were separated by electrophoresis on 10% SDS-PAGE gels and transferred onto nitrocellulose membranes. .. The blots were blocked with 1% skimmed milk and probed overnight with the primary antibodies: Heme Oxygenase 1 (Santa Cruz Biotechnology, sc-10789) polyclonal antibody, NQO1 (Santa Cruz Biotechnology, sc-271116) monoclonal antibody, or β-actin (Santa Cruz Biotechnology, sc-130657) polyclonal antibody.

Incubation:

Article Title: Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model
Article Snippet: .. The latter were blocked with 5% milk, followed by incubation with the following antibodies: TNF-α , PGC-1α (Abcam, Cambridge, MA), PAI-1 (BD Bioscience, San Jose, CA), 3-NT (Millipore, Billerica, MA), 4-HNE (Alpha Diagnostic International, San Antonio, TX), HK II, β -actin, and NQO1 (SantaCruz Biotechnology, Santa Cruz, CA). .. After those membranes were washed with Tris-buffered saline (pH 7.2) containing 0.05% Tween 20 and incubated with the appropriate secondary antibodies.

Article Title: Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways
Article Snippet: .. Following the transfer, membranes were blocked with TBST (10 mmol/l Tris-HCl pH 7.4, 150 mmol/l NaCl, and 0.1% Tween 20) containing 5% nonfat dry milk (blocking buffer) and incubated with the primary antibodies (diluted in blocking buffer overnight at 4°C) against SIRT-1 (Cell Signaling, cat. #9475), p-SIRT-1 (Cell Signaling, cat. #2314), Akt (Cell Signaling, cat. #9272), p-Akt (Cell Signaling, cat. #9271), AMPKα (Cell Signaling, cat. #5831), p-AMPKα (Cell Signaling, cat. #2535), NQO1 (Santa Cruz, cat. #sc-16464), β-actin (Cell Signaling, cat. #4970), and β-tubulin (Cell Signaling, cat. #2146). .. Membranes were incubated with goat anti-rabbit immunoglobulin (IgG) secondary antibody (Santa Cruz, cat. #sc-2030) for 1 h at room temperature, and washed 5 times.

Article Title: Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas
Article Snippet: Blots were incubated with 5% (w/v) nonfat dry milk in PBS with 0.05% (w/v) Tween 20 (Millipore Sigma, P9416) (PBST) at room temp for 1 hr to block nonspecific binding, and overnight at 4°C with primary antibodies in 3% BSA (w/v) in PBST and finally with HRP-conjugated secondary antibody in blocking buffer. .. The following antibodies were used: PCNA (BD Pharmingen, #555566), cyclin D1 (Santa Cruz, sc-8396), pERK (Cell Signaling, #9101), ERK (Cell Signaling, #9102), p62 (Progen, GP62-C), LC3B (Cell Signaling, #2775), Keap1 (Cell Signaling D6B12), NRF2 (Santa Cruz, sc-722), NQO1 (Santa Cruz, 271116), tubulin (Sigma, T9026), Hes1 (Santa Cruz, sc-25392), CK19 (Santa Cruz, sc-33111), p27 (Santa Cruz, sc-528), p53 (Cell Signaling, 2524), Sox9 (Santa Cruz, sc-20095), Ki67 (GeneTex, GTX 16667), MDM2 (Santa Cruz, sc-965), ALDH (Abcam, ab24343), p-Histone H2A.X (Ser 139) (Santa Cruz, sc-517348).

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
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Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization. .. Subsequently, the membranes were washed and incubated at room temperature with a horseradish peroxidase-conjugated secondary polyclonal anti-rabbit-IgG antibody (Sigma-Aldrich Cat# A0545, RRID:AB_257896), anti-mouse-IgG antibody (Abcam Cat# ab6728, RRID:AB_955440) or anti-goat-IgG antibody (Santa Cruz Biotechnology Cat# sc-2020, RRID:AB_631728), respectively, for 1.5 h. The membranes used for detection of p-eIF2α were stripped for 30 min with mild stripping buffer following washing and incubation with anti-eIF2α antibody to detect total eIF2α protein expression on the same membranes.

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
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Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: .. Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology). .. Membranes were then exposed to an anti-rabbit secondary antibody conjugated to horseradish peroxidase (Cell Signaling Technology) for 1 h at room temperature.

Article Title: PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy
Article Snippet: The blots were blocked with 1% skimmed milk and probed overnight with the primary antibodies: Heme Oxygenase 1 (Santa Cruz Biotechnology, sc-10789) polyclonal antibody, NQO1 (Santa Cruz Biotechnology, sc-271116) monoclonal antibody, or β-actin (Santa Cruz Biotechnology, sc-130657) polyclonal antibody. .. Following a 2 hr incubation period with peroxidase-conjugated secondary antibodies, proteins were detected using enhanced chemiluminescence (Fisher Scientific, 12316992).

Article Title: Sulforaphane Inhibits Mitochondrial Permeability Transition and Oxidative Stress
Article Snippet: .. Equal amounts of mitochondrial protein from each sample were separated by SDS-PAGE (4–12% Bis-Tris gels) (Invitrogen) and transferred to PVDF membranes (Invitrogen), and then incubated with primary antibodies (overnight at 4°C) with NQO1 1:500 (Santa Cruz), Cyclophilin D 1:30000 (Mitosciences), Glutathione Peroxidase 1:500 (Abcam), Thioredoxin 2 1:150000 (Santa Cruz), Malic Enzyme 3 1:5000 (Sigma), Isocitrate Dehydrogenase 2 1:2000 (Santa Cruz), Manganese Superoxide Dismutase 1:250 (Santa Cruz), GAPDH 1:500000 (Abcam), VDAC 1:50,000–1:300,000 (Mitosciences). .. The membranes were then washed with PBST and incubated for 1 h at RT in HRP conjugated antibodies at 1:2000 dilution for 1 hr at RT.

Stripping Membranes:

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization. .. Subsequently, the membranes were washed and incubated at room temperature with a horseradish peroxidase-conjugated secondary polyclonal anti-rabbit-IgG antibody (Sigma-Aldrich Cat# A0545, RRID:AB_257896), anti-mouse-IgG antibody (Abcam Cat# ab6728, RRID:AB_955440) or anti-goat-IgG antibody (Santa Cruz Biotechnology Cat# sc-2020, RRID:AB_631728), respectively, for 1.5 h. The membranes used for detection of p-eIF2α were stripped for 30 min with mild stripping buffer following washing and incubation with anti-eIF2α antibody to detect total eIF2α protein expression on the same membranes.

Activity Assay:

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization. .. The HRP activity was detected using chemiluminescent reagents (Amersham ECL Select Western Blotting Detection Reagent, GE healthcare, Freiburg, Germany).

Expressing:

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization. .. Subsequently, the membranes were washed and incubated at room temperature with a horseradish peroxidase-conjugated secondary polyclonal anti-rabbit-IgG antibody (Sigma-Aldrich Cat# A0545, RRID:AB_257896), anti-mouse-IgG antibody (Abcam Cat# ab6728, RRID:AB_955440) or anti-goat-IgG antibody (Santa Cruz Biotechnology Cat# sc-2020, RRID:AB_631728), respectively, for 1.5 h. The membranes used for detection of p-eIF2α were stripped for 30 min with mild stripping buffer following washing and incubation with anti-eIF2α antibody to detect total eIF2α protein expression on the same membranes.

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: Western Blotting In order to determine the hepatic protein expression of PPARγ, NQO1, and HO-1, total protein was isolated from the liver in a cold radio-immunoprecipitation assay (RIPA) lysis buffer (Beyotime Institute of Biotechnology, Nantong, Jiangsu, China) with 1% phosphatase inhibitor cocktail and 1% phenylmethanesulfonyl fluoride (PMSF). .. The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight.

BIA-KA:

Article Title: Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways
Article Snippet: Protein content was determined using a BCA Protein Assay Kit (Pierce, Rockford, IL) and SDS samples were prepared. .. Following the transfer, membranes were blocked with TBST (10 mmol/l Tris-HCl pH 7.4, 150 mmol/l NaCl, and 0.1% Tween 20) containing 5% nonfat dry milk (blocking buffer) and incubated with the primary antibodies (diluted in blocking buffer overnight at 4°C) against SIRT-1 (Cell Signaling, cat. #9475), p-SIRT-1 (Cell Signaling, cat. #2314), Akt (Cell Signaling, cat. #9272), p-Akt (Cell Signaling, cat. #9271), AMPKα (Cell Signaling, cat. #5831), p-AMPKα (Cell Signaling, cat. #2535), NQO1 (Santa Cruz, cat. #sc-16464), β-actin (Cell Signaling, cat. #4970), and β-tubulin (Cell Signaling, cat. #2146).

Modification:

Article Title: Corynoline Isolated from Corydalis bungeana Turcz. Exhibits Anti-Inflammatory Effects via Modulation of Nfr2 and MAPKs
Article Snippet: Dulbecco’s Modified Eagle’s Medium (DMEM) and fetal bovine serum (FBS) were obtained from Invitrogen-Gibco (Grand Island, NY, USA). .. The anti-COX2, anti-iNOS, anti-p38, anti-ERK, anti-JNK, anti-actin, anti-HO1, anti-NQO1 and anti-Akt primary antibodies and secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Western Blot:

Article Title: Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model
Article Snippet: Paragraph title: 4.5. Western Blot Assay ... The latter were blocked with 5% milk, followed by incubation with the following antibodies: TNF-α , PGC-1α (Abcam, Cambridge, MA), PAI-1 (BD Bioscience, San Jose, CA), 3-NT (Millipore, Billerica, MA), 4-HNE (Alpha Diagnostic International, San Antonio, TX), HK II, β -actin, and NQO1 (SantaCruz Biotechnology, Santa Cruz, CA).

Article Title: Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways
Article Snippet: Paragraph title: Western blot analysis ... Following the transfer, membranes were blocked with TBST (10 mmol/l Tris-HCl pH 7.4, 150 mmol/l NaCl, and 0.1% Tween 20) containing 5% nonfat dry milk (blocking buffer) and incubated with the primary antibodies (diluted in blocking buffer overnight at 4°C) against SIRT-1 (Cell Signaling, cat. #9475), p-SIRT-1 (Cell Signaling, cat. #2314), Akt (Cell Signaling, cat. #9272), p-Akt (Cell Signaling, cat. #9271), AMPKα (Cell Signaling, cat. #5831), p-AMPKα (Cell Signaling, cat. #2535), NQO1 (Santa Cruz, cat. #sc-16464), β-actin (Cell Signaling, cat. #4970), and β-tubulin (Cell Signaling, cat. #2146).

Article Title: Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas
Article Snippet: Blots were developed using enhanced chemiluminescence detection kit (Western Lighting Plus-ECL kit, PerkinElmer, NEL103001EA). .. The following antibodies were used: PCNA (BD Pharmingen, #555566), cyclin D1 (Santa Cruz, sc-8396), pERK (Cell Signaling, #9101), ERK (Cell Signaling, #9102), p62 (Progen, GP62-C), LC3B (Cell Signaling, #2775), Keap1 (Cell Signaling D6B12), NRF2 (Santa Cruz, sc-722), NQO1 (Santa Cruz, 271116), tubulin (Sigma, T9026), Hes1 (Santa Cruz, sc-25392), CK19 (Santa Cruz, sc-33111), p27 (Santa Cruz, sc-528), p53 (Cell Signaling, 2524), Sox9 (Santa Cruz, sc-20095), Ki67 (GeneTex, GTX 16667), MDM2 (Santa Cruz, sc-965), ALDH (Abcam, ab24343), p-Histone H2A.X (Ser 139) (Santa Cruz, sc-517348).

Article Title: Delayed treatment with oleanolic acid attenuates tubulointerstitial fibrosis in chronic cyclosporine nephropathy through Nrf2/HO-1 signaling
Article Snippet: Paragraph title: Western blotting ... Specifically, proteins were separated by SDS-PAGE, transferred to nitrocellulose membranes, and detected with the following antibody concentrations: Nrf2 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), Keap1 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), HO-1 (1:1000; BD Biosciences, California, USA), NQO1 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), Bcl-2 (1:500; Santa Cruz Biotechnology Inc, Texas, USA), Bax (1:500; Santa Cruz Biotechnology Inc, Texas, USA), SOD1 (1:5000; Assay Designs, MI, USA), SOD2 (1:10000; Abcam, Cambridge, UK), Catalase (1:2000; Abcam, Cambridge, UK), and β-actin (1:10000; Sigma-Aldrich, MO, USA).

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: .. After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific). .. Immunoblot imaging was captured by using a BioSpectrum AC Imaging System (Ultra-Violet Products).

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization. .. The HRP activity was detected using chemiluminescent reagents (Amersham ECL Select Western Blotting Detection Reagent, GE healthcare, Freiburg, Germany).

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: Paragraph title: 2.9. Western Blotting ... The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight.

Article Title: Scolopendra subspinipes mutilans attenuates neuroinflammation in symptomatic hSOD1G93A mice
Article Snippet: .. The primary antibodies employed for Western blotting and immunohistochemistry were as follows: anti-Iba-1 (diluted 1:1,000, Wako, Japan), anti-GFAP (diluted 1:3,000, Millipore Corp., MA, USA), anti-MAP2 (diluted 1:500, Millipore Corp., MA, USA), anti-HO1 (diluted 1:1,000, Abcam, MA, USA), anti-NQO1 (diluted 1:1,000, Santa Cruz Biotechnology, CA, USA), anti-human SOD1 (diluted 1:2,000, Calbiochem, CA, USA), and anti-CD14 (diluted 1:500, BD Biosciences, CA, USA). .. Anti-α-tubulin (diluted 1:5,000, Abcam, MA, USA) was used as a loading control.

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: Paragraph title: Western blot analysis ... Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology).

Article Title: PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy
Article Snippet: Paragraph title: Western Blotting ... The blots were blocked with 1% skimmed milk and probed overnight with the primary antibodies: Heme Oxygenase 1 (Santa Cruz Biotechnology, sc-10789) polyclonal antibody, NQO1 (Santa Cruz Biotechnology, sc-271116) monoclonal antibody, or β-actin (Santa Cruz Biotechnology, sc-130657) polyclonal antibody.

Translocation Assay:

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: To determine the nuclear translocation of Nrf2, the supernatants from the first step were gathered and re-centrifuged. .. The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight.

Immunohistochemistry:

Article Title: Scolopendra subspinipes mutilans attenuates neuroinflammation in symptomatic hSOD1G93A mice
Article Snippet: .. The primary antibodies employed for Western blotting and immunohistochemistry were as follows: anti-Iba-1 (diluted 1:1,000, Wako, Japan), anti-GFAP (diluted 1:3,000, Millipore Corp., MA, USA), anti-MAP2 (diluted 1:500, Millipore Corp., MA, USA), anti-HO1 (diluted 1:1,000, Abcam, MA, USA), anti-NQO1 (diluted 1:1,000, Santa Cruz Biotechnology, CA, USA), anti-human SOD1 (diluted 1:2,000, Calbiochem, CA, USA), and anti-CD14 (diluted 1:500, BD Biosciences, CA, USA). .. Anti-α-tubulin (diluted 1:5,000, Abcam, MA, USA) was used as a loading control.

Imaging:

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific). .. Immunoblot imaging was captured by using a BioSpectrum AC Imaging System (Ultra-Violet Products).

Protein Concentration:

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: Protein concentration was determined using Bradford protein assay kit (BioRad). .. After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific).

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: Immunoblotting Preparation of liver homogenates, determination of the protein concentration in the homogenates, protein separation by 12.5% SDS-PAGE, and transfer to a nitrocellulose membrane were carried out as recently described [ ]. .. After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization.

Sonication:

Article Title: Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2
Article Snippet: The antibodies for Keap1, Nrf2, α-tubulin, β-actin, NQO1, HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA). .. Following sonication, cell lysates were electrophoresed through sodium dodecyl sulfate–polyacrylamide gel and subjected to immunoblot analysis.

Recombinant:

Article Title: PAR-1 is a novel mechano-sensor transducing laminar flow-mediated endothelial signaling
Article Snippet: Reagents and antibodies Recombinant human TNF-α was purchased from R & D Systems (Wiesbaden-Norderstedt, Germany). .. Anti-PAR-1 (ATAP2), anti-NQO1, anti-eNOS, and anti-VCAM-1 were from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and anti-phospho-HDAC5, anti-HDAC5, anti-phospho-ERK5, anti-ERK5, anti-phospho-AKT, anti-AKT, anti-phospho-AMPK, anti-AMPK, anti-phospho-eNOS, anti-EEA1, anti-phospho-Src, anti-phospho-FAK, anti-phospho-Erk1/2, anti-Erk1/2 and anti-VE-cadherin were from Cell Signaling Technology (Danvers, MA, USA).

Fluorescence:

Article Title: Scolopendra subspinipes mutilans attenuates neuroinflammation in symptomatic hSOD1G93A mice
Article Snippet: An avidin-biotin peroxidase complex (ABC) kit, 3, 3′-diaminobenzidine tetrahydrochloride (DAB), and mounting medium for performing fluorescence analysis with DAPI were procured from Vector Laboratories (Burlingame, CA, USA). .. The primary antibodies employed for Western blotting and immunohistochemistry were as follows: anti-Iba-1 (diluted 1:1,000, Wako, Japan), anti-GFAP (diluted 1:3,000, Millipore Corp., MA, USA), anti-MAP2 (diluted 1:500, Millipore Corp., MA, USA), anti-HO1 (diluted 1:1,000, Abcam, MA, USA), anti-NQO1 (diluted 1:1,000, Santa Cruz Biotechnology, CA, USA), anti-human SOD1 (diluted 1:2,000, Calbiochem, CA, USA), and anti-CD14 (diluted 1:500, BD Biosciences, CA, USA).

Isolation:

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: Western Blotting In order to determine the hepatic protein expression of PPARγ, NQO1, and HO-1, total protein was isolated from the liver in a cold radio-immunoprecipitation assay (RIPA) lysis buffer (Beyotime Institute of Biotechnology, Nantong, Jiangsu, China) with 1% phosphatase inhibitor cocktail and 1% phenylmethanesulfonyl fluoride (PMSF). .. The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight.

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: Western blot analysis Total protein was isolated from liver tissues by homogenization in an ice-cold lysis buffer containing 20 mM HEPES-KOH (pH 7.9), 125 mM NaCl, 10% glycerol, 0.3% Triton X-100, 1 mM EDTA, 0.5% NP-40, 10 mM β-phosphoglycerate, 1 mM Na3 VO4 , 5 mM NaF, 1 mM aprotinin, 1 mM phenylmethanesulfonylfluoride, and 1 mM leupeptin. .. Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology).

Article Title: Sulforaphane Inhibits Mitochondrial Permeability Transition and Oxidative Stress
Article Snippet: Freshly Isolated liver mitochondria and liver homogenates were lysed in RIPA buffer containing a cocktail of protease and phosphatase inhibitors (Calbiochem). .. Equal amounts of mitochondrial protein from each sample were separated by SDS-PAGE (4–12% Bis-Tris gels) (Invitrogen) and transferred to PVDF membranes (Invitrogen), and then incubated with primary antibodies (overnight at 4°C) with NQO1 1:500 (Santa Cruz), Cyclophilin D 1:30000 (Mitosciences), Glutathione Peroxidase 1:500 (Abcam), Thioredoxin 2 1:150000 (Santa Cruz), Malic Enzyme 3 1:5000 (Sigma), Isocitrate Dehydrogenase 2 1:2000 (Santa Cruz), Manganese Superoxide Dismutase 1:250 (Santa Cruz), GAPDH 1:500000 (Abcam), VDAC 1:50,000–1:300,000 (Mitosciences).

Bicinchoninic Acid Protein Assay:

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: Protein concentrations were determined by the bicinchoninic acid protein assay kit (Interchim, Montluçon, France) with BSA as standard. .. After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization.

Transfection:

Article Title: Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2
Article Snippet: Paragraph title: Transient transfection of small interfering RNA, antibodies and immunoblot analysis ... The antibodies for Keap1, Nrf2, α-tubulin, β-actin, NQO1, HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA).

Avidin-Biotin Assay:

Article Title: Scolopendra subspinipes mutilans attenuates neuroinflammation in symptomatic hSOD1G93A mice
Article Snippet: An avidin-biotin peroxidase complex (ABC) kit, 3, 3′-diaminobenzidine tetrahydrochloride (DAB), and mounting medium for performing fluorescence analysis with DAPI were procured from Vector Laboratories (Burlingame, CA, USA). .. The primary antibodies employed for Western blotting and immunohistochemistry were as follows: anti-Iba-1 (diluted 1:1,000, Wako, Japan), anti-GFAP (diluted 1:3,000, Millipore Corp., MA, USA), anti-MAP2 (diluted 1:500, Millipore Corp., MA, USA), anti-HO1 (diluted 1:1,000, Abcam, MA, USA), anti-NQO1 (diluted 1:1,000, Santa Cruz Biotechnology, CA, USA), anti-human SOD1 (diluted 1:2,000, Calbiochem, CA, USA), and anti-CD14 (diluted 1:500, BD Biosciences, CA, USA).

Protein Extraction:

Article Title: Delayed treatment with oleanolic acid attenuates tubulointerstitial fibrosis in chronic cyclosporine nephropathy through Nrf2/HO-1 signaling
Article Snippet: Western blotting For Western blot analysis, total protein of renal cortical tissues was extracted with a Pro-Prep Protein Extraction Solution (Intron Biotechnology, Gyeonggi-do, Korea) according to the manufacturer’s instructions. .. Specifically, proteins were separated by SDS-PAGE, transferred to nitrocellulose membranes, and detected with the following antibody concentrations: Nrf2 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), Keap1 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), HO-1 (1:1000; BD Biosciences, California, USA), NQO1 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), Bcl-2 (1:500; Santa Cruz Biotechnology Inc, Texas, USA), Bax (1:500; Santa Cruz Biotechnology Inc, Texas, USA), SOD1 (1:5000; Assay Designs, MI, USA), SOD2 (1:10000; Abcam, Cambridge, UK), Catalase (1:2000; Abcam, Cambridge, UK), and β-actin (1:10000; Sigma-Aldrich, MO, USA).

Bradford Protein Assay:

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: Protein concentration was determined using Bradford protein assay kit (BioRad). .. After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific).

Staining:

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: Reversible Ponceau S (Carl Roth, Karlsruhe, Germany) staining was performed to check equal protein transfer to the membranes. .. After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization.

SDS Page:

Article Title: Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model
Article Snippet: Briefly, protein was separated on 10% SDS-PAGE gels and transferred to nitrocellulose membranes (Bio-Rad, Hercules, CA). .. The latter were blocked with 5% milk, followed by incubation with the following antibodies: TNF-α , PGC-1α (Abcam, Cambridge, MA), PAI-1 (BD Bioscience, San Jose, CA), 3-NT (Millipore, Billerica, MA), 4-HNE (Alpha Diagnostic International, San Antonio, TX), HK II, β -actin, and NQO1 (SantaCruz Biotechnology, Santa Cruz, CA).

Article Title: Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas
Article Snippet: IB analysis was performed on tissue or cell lysates that were separated by SDS-PAGE and transferred to nitrocellulose membranes. .. The following antibodies were used: PCNA (BD Pharmingen, #555566), cyclin D1 (Santa Cruz, sc-8396), pERK (Cell Signaling, #9101), ERK (Cell Signaling, #9102), p62 (Progen, GP62-C), LC3B (Cell Signaling, #2775), Keap1 (Cell Signaling D6B12), NRF2 (Santa Cruz, sc-722), NQO1 (Santa Cruz, 271116), tubulin (Sigma, T9026), Hes1 (Santa Cruz, sc-25392), CK19 (Santa Cruz, sc-33111), p27 (Santa Cruz, sc-528), p53 (Cell Signaling, 2524), Sox9 (Santa Cruz, sc-20095), Ki67 (GeneTex, GTX 16667), MDM2 (Santa Cruz, sc-965), ALDH (Abcam, ab24343), p-Histone H2A.X (Ser 139) (Santa Cruz, sc-517348).

Article Title: Delayed treatment with oleanolic acid attenuates tubulointerstitial fibrosis in chronic cyclosporine nephropathy through Nrf2/HO-1 signaling
Article Snippet: .. Specifically, proteins were separated by SDS-PAGE, transferred to nitrocellulose membranes, and detected with the following antibody concentrations: Nrf2 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), Keap1 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), HO-1 (1:1000; BD Biosciences, California, USA), NQO1 (1:1000; Santa Cruz Biotechnology Inc, Texas, USA), Bcl-2 (1:500; Santa Cruz Biotechnology Inc, Texas, USA), Bax (1:500; Santa Cruz Biotechnology Inc, Texas, USA), SOD1 (1:5000; Assay Designs, MI, USA), SOD2 (1:10000; Abcam, Cambridge, UK), Catalase (1:2000; Abcam, Cambridge, UK), and β-actin (1:10000; Sigma-Aldrich, MO, USA). ..

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: 15 μg protein were separated by 10% SDS-PAGE and electro-transferred to nitrocellulose membrane (Pall Corporation, Pensacola, FL, USA). .. After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization.

Article Title: PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy
Article Snippet: Proteins were separated by electrophoresis on 10% SDS-PAGE gels and transferred onto nitrocellulose membranes. .. The blots were blocked with 1% skimmed milk and probed overnight with the primary antibodies: Heme Oxygenase 1 (Santa Cruz Biotechnology, sc-10789) polyclonal antibody, NQO1 (Santa Cruz Biotechnology, sc-271116) monoclonal antibody, or β-actin (Santa Cruz Biotechnology, sc-130657) polyclonal antibody.

Article Title: Sulforaphane Inhibits Mitochondrial Permeability Transition and Oxidative Stress
Article Snippet: .. Equal amounts of mitochondrial protein from each sample were separated by SDS-PAGE (4–12% Bis-Tris gels) (Invitrogen) and transferred to PVDF membranes (Invitrogen), and then incubated with primary antibodies (overnight at 4°C) with NQO1 1:500 (Santa Cruz), Cyclophilin D 1:30000 (Mitosciences), Glutathione Peroxidase 1:500 (Abcam), Thioredoxin 2 1:150000 (Santa Cruz), Malic Enzyme 3 1:5000 (Sigma), Isocitrate Dehydrogenase 2 1:2000 (Santa Cruz), Manganese Superoxide Dismutase 1:250 (Santa Cruz), GAPDH 1:500000 (Abcam), VDAC 1:50,000–1:300,000 (Mitosciences). .. The membranes were then washed with PBST and incubated for 1 h at RT in HRP conjugated antibodies at 1:2000 dilution for 1 hr at RT.

Software:

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific). .. The protein bands were quantified using ImageQuant software (GE-Healthcare) and digitally converted for statistical analysis.

Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil
Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization. .. The signal intensities of specific bands were detected with a Bio-Imaging system (Syngene, Cambridge, UK) and quantified using Syngene GeneTools software.

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight. .. The protein quantity was determined by densitometry analysis using ImageJ software (version 1.47, National Institutes of Health, Bethesda, MD, USA).

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology). .. Densitometry analysis was performed using ImageJ software (National Institutes of Health, Bethesda, MD, USA).

Article Title: PMI: A ΔΨm Independent Pharmacological Regulator of Mitophagy
Article Snippet: Immunoreactive bands were analyzed by performing densitometry with ImageJ software. .. The blots were blocked with 1% skimmed milk and probed overnight with the primary antibodies: Heme Oxygenase 1 (Santa Cruz Biotechnology, sc-10789) polyclonal antibody, NQO1 (Santa Cruz Biotechnology, sc-271116) monoclonal antibody, or β-actin (Santa Cruz Biotechnology, sc-130657) polyclonal antibody.

Article Title: Sulforaphane Inhibits Mitochondrial Permeability Transition and Oxidative Stress
Article Snippet: Equal amounts of mitochondrial protein from each sample were separated by SDS-PAGE (4–12% Bis-Tris gels) (Invitrogen) and transferred to PVDF membranes (Invitrogen), and then incubated with primary antibodies (overnight at 4°C) with NQO1 1:500 (Santa Cruz), Cyclophilin D 1:30000 (Mitosciences), Glutathione Peroxidase 1:500 (Abcam), Thioredoxin 2 1:150000 (Santa Cruz), Malic Enzyme 3 1:5000 (Sigma), Isocitrate Dehydrogenase 2 1:2000 (Santa Cruz), Manganese Superoxide Dismutase 1:250 (Santa Cruz), GAPDH 1:500000 (Abcam), VDAC 1:50,000–1:300,000 (Mitosciences). .. Densitometric analysis of the protein bands was performed using the Image J software.

Binding Assay:

Article Title: Stress Activated NRF2-MDM2 Cascade Controls Neoplastic Progression in Pancreas
Article Snippet: Blots were incubated with 5% (w/v) nonfat dry milk in PBS with 0.05% (w/v) Tween 20 (Millipore Sigma, P9416) (PBST) at room temp for 1 hr to block nonspecific binding, and overnight at 4°C with primary antibodies in 3% BSA (w/v) in PBST and finally with HRP-conjugated secondary antibody in blocking buffer. .. The following antibodies were used: PCNA (BD Pharmingen, #555566), cyclin D1 (Santa Cruz, sc-8396), pERK (Cell Signaling, #9101), ERK (Cell Signaling, #9102), p62 (Progen, GP62-C), LC3B (Cell Signaling, #2775), Keap1 (Cell Signaling D6B12), NRF2 (Santa Cruz, sc-722), NQO1 (Santa Cruz, 271116), tubulin (Sigma, T9026), Hes1 (Santa Cruz, sc-25392), CK19 (Santa Cruz, sc-33111), p27 (Santa Cruz, sc-528), p53 (Cell Signaling, 2524), Sox9 (Santa Cruz, sc-20095), Ki67 (GeneTex, GTX 16667), MDM2 (Santa Cruz, sc-965), ALDH (Abcam, ab24343), p-Histone H2A.X (Ser 139) (Santa Cruz, sc-517348).

Radio Immunoprecipitation:

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: Western Blotting In order to determine the hepatic protein expression of PPARγ, NQO1, and HO-1, total protein was isolated from the liver in a cold radio-immunoprecipitation assay (RIPA) lysis buffer (Beyotime Institute of Biotechnology, Nantong, Jiangsu, China) with 1% phosphatase inhibitor cocktail and 1% phenylmethanesulfonyl fluoride (PMSF). .. The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight.

Homogenization:

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: Western blot analysis Total protein was isolated from liver tissues by homogenization in an ice-cold lysis buffer containing 20 mM HEPES-KOH (pH 7.9), 125 mM NaCl, 10% glycerol, 0.3% Triton X-100, 1 mM EDTA, 0.5% NP-40, 10 mM β-phosphoglycerate, 1 mM Na3 VO4 , 5 mM NaF, 1 mM aprotinin, 1 mM phenylmethanesulfonylfluoride, and 1 mM leupeptin. .. Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology).

Small Interfering RNA:

Article Title: Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2
Article Snippet: Paragraph title: Transient transfection of small interfering RNA, antibodies and immunoblot analysis ... The antibodies for Keap1, Nrf2, α-tubulin, β-actin, NQO1, HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA).

Migration:

Article Title: Extendin-4 protects kidney from acute ischemia-reperfusion injury through upregulation of NRF2 signaling
Article Snippet: After protein migration, the proteins were blotted on PVDF membrane (Immobilon 0.45 m, Millipore). .. After blocking, membranes were incubated with first antibodies [total Akt (1:1000, Cell Signaling), phosphorylated (p)-Akt (1:1000, Cell Signaling), NOX-1 (1:1500, Sigma), NOX-2 (1:750, Sigma), Nrf2 (1:200; Santa Cruz Biotechnology) and NQO1 (1:200; Santa Cruz Biotechnology) for 2 h. Subsequently, the membranes were incubated with the desired secondary antibody for another 2 h. Immuoreactive signals were detected by incubation with horseradish peroxidase conjugated-secondary followed by enhanced chemiluminescent detection using Pierce ECL Western Blotting Substrate (Thermo Scientific).

Lysis:

Article Title: Thymoquinone ameliorates diabetic phenotype in Diet-Induced Obesity mice via activation of SIRT-1-dependent pathways
Article Snippet: After exposure, HepG2 cells were solubilized in RIPA lysis buffer. .. Following the transfer, membranes were blocked with TBST (10 mmol/l Tris-HCl pH 7.4, 150 mmol/l NaCl, and 0.1% Tween 20) containing 5% nonfat dry milk (blocking buffer) and incubated with the primary antibodies (diluted in blocking buffer overnight at 4°C) against SIRT-1 (Cell Signaling, cat. #9475), p-SIRT-1 (Cell Signaling, cat. #2314), Akt (Cell Signaling, cat. #9272), p-Akt (Cell Signaling, cat. #9271), AMPKα (Cell Signaling, cat. #5831), p-AMPKα (Cell Signaling, cat. #2535), NQO1 (Santa Cruz, cat. #sc-16464), β-actin (Cell Signaling, cat. #4970), and β-tubulin (Cell Signaling, cat. #2146).

Article Title: Regressive Effect of Myricetin on Hepatic Steatosis in Mice Fed a High-Fat Diet
Article Snippet: Western Blotting In order to determine the hepatic protein expression of PPARγ, NQO1, and HO-1, total protein was isolated from the liver in a cold radio-immunoprecipitation assay (RIPA) lysis buffer (Beyotime Institute of Biotechnology, Nantong, Jiangsu, China) with 1% phosphatase inhibitor cocktail and 1% phenylmethanesulfonyl fluoride (PMSF). .. The membranes were blocked in Tris-buffered saline (TBS) containing 5% (w/v ) BSA and thereafter incubated with the primary antibodies, including Nrf2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PPARγ (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and Lamin B1 (Serotec Ltd., Oxford, UK) at 4 °C overnight.

Article Title: CB1 receptor blockade ameliorates hepatic fat infiltration and inflammation and increases Nrf2-AMPK pathway in a rat model of severely uncontrolled diabetes
Article Snippet: Western blot analysis Total protein was isolated from liver tissues by homogenization in an ice-cold lysis buffer containing 20 mM HEPES-KOH (pH 7.9), 125 mM NaCl, 10% glycerol, 0.3% Triton X-100, 1 mM EDTA, 0.5% NP-40, 10 mM β-phosphoglycerate, 1 mM Na3 VO4 , 5 mM NaF, 1 mM aprotinin, 1 mM phenylmethanesulfonylfluoride, and 1 mM leupeptin. .. Membranes were incubated overnight at 4°C with the following primary antibodies: anti-β actin (AbClon, Seoul, South Korea), anti-HO-1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), NQO1 (Santa Cruz Biotechnology), anti-phospho AMPK (Cell Signaling Technology, Danvers, MA, USA), and anti-AMPK (Cell Signaling Technology).

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  • 99
    Santa Cruz Biotechnology anti nqo1
    eIF4GI and eIF4GII, but not DAP5, are degraded under oxidative stress. (A) NIH-3T3 cells were untreated or treated with increasing concentration of H 2 O 2 in the presence or absence of lactacystin (described in Alard et al., 2009 ), and protein extracts were subjected to western-blotting as indicated. (B) NIH-3T3 cell extracts were subjected to western-blotting with the indicated antibodies either directly (input) or after immunoprecipitation (IP) with either eIF4GI or eIF4GII antibodies (left). NIH-3T3 extracts of cells either untransfected of transfected with <t>NQO1</t> cDNA were subjected to western-blotting with the indicated antibodies either directly (input) or after immunoprecipitation (IP) with NQO1 antibodies (right). (C) NIH-3T3 cells were untreated or treated with 300 μM dicumarol (Dic) at different times and proteins were visualized by western-blotting as indicated. (D) Following transfection with HA-tagged, full-length or N-terminal cDNAs, NIH-3T3 cells were untreated or treated with 300 μM dicumarol for 8 h and proteins visualized by western-blotting as indicated.
    Anti Nqo1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti nqo1/product/Santa Cruz Biotechnology
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    anti nqo1 - by Bioz Stars, 2020-02
    99/100 stars
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    86
    Santa Cruz Biotechnology mouse anti nqo1
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
    Mouse Anti Nqo1, 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/mouse anti nqo1/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti nqo1 - by Bioz Stars, 2020-02
    86/100 stars
      Buy from Supplier

    Image Search Results


    eIF4GI and eIF4GII, but not DAP5, are degraded under oxidative stress. (A) NIH-3T3 cells were untreated or treated with increasing concentration of H 2 O 2 in the presence or absence of lactacystin (described in Alard et al., 2009 ), and protein extracts were subjected to western-blotting as indicated. (B) NIH-3T3 cell extracts were subjected to western-blotting with the indicated antibodies either directly (input) or after immunoprecipitation (IP) with either eIF4GI or eIF4GII antibodies (left). NIH-3T3 extracts of cells either untransfected of transfected with NQO1 cDNA were subjected to western-blotting with the indicated antibodies either directly (input) or after immunoprecipitation (IP) with NQO1 antibodies (right). (C) NIH-3T3 cells were untreated or treated with 300 μM dicumarol (Dic) at different times and proteins were visualized by western-blotting as indicated. (D) Following transfection with HA-tagged, full-length or N-terminal cDNAs, NIH-3T3 cells were untreated or treated with 300 μM dicumarol for 8 h and proteins visualized by western-blotting as indicated.

    Journal: Frontiers in Genetics

    Article Title: Differential Regulation of the Three Eukaryotic mRNA Translation Initiation Factor (eIF) 4Gs by the Proteasome

    doi: 10.3389/fgene.2019.00254

    Figure Lengend Snippet: eIF4GI and eIF4GII, but not DAP5, are degraded under oxidative stress. (A) NIH-3T3 cells were untreated or treated with increasing concentration of H 2 O 2 in the presence or absence of lactacystin (described in Alard et al., 2009 ), and protein extracts were subjected to western-blotting as indicated. (B) NIH-3T3 cell extracts were subjected to western-blotting with the indicated antibodies either directly (input) or after immunoprecipitation (IP) with either eIF4GI or eIF4GII antibodies (left). NIH-3T3 extracts of cells either untransfected of transfected with NQO1 cDNA were subjected to western-blotting with the indicated antibodies either directly (input) or after immunoprecipitation (IP) with NQO1 antibodies (right). (C) NIH-3T3 cells were untreated or treated with 300 μM dicumarol (Dic) at different times and proteins were visualized by western-blotting as indicated. (D) Following transfection with HA-tagged, full-length or N-terminal cDNAs, NIH-3T3 cells were untreated or treated with 300 μM dicumarol for 8 h and proteins visualized by western-blotting as indicated.

    Article Snippet: The antibodies used were as follows: anti-eIF4GI and anti-eIF4GII (gifts of Prof. Nahum Sonenberg); anti-DAP5 (CliniSciences #610742); anti-HA-7 (Sigma); anti-β-tubulin (GeneTex #6288022); anti-4E-BP1, anti-NRF2 and anti-p53 (Cell Signaling Technologies #9452, #12721, and #1C12, respectively); anti-Core 20S (Enzo Life Sciences #PW8155); and anti-NQO1 (Santa Cruz #C19).

    Techniques: Concentration Assay, Western Blot, Immunoprecipitation, Transfection

    Induction of NRF2 and NQO1 proteins under oxidative stress is independent of DAP5. Protein extracts of stably transfected NIH-3T3 cells grown in the absence or presence of doxycycline (Dox) for 48 h and untreated or treated with 1 mM H 2 O 2 for 4 h were subjected to western-blotting with the indicated antibodies. The bottom-to-top α–β–γ symbols denote hypo- to hyperphosphorylated 4E-BP1 isoforms.

    Journal: Frontiers in Genetics

    Article Title: Differential Regulation of the Three Eukaryotic mRNA Translation Initiation Factor (eIF) 4Gs by the Proteasome

    doi: 10.3389/fgene.2019.00254

    Figure Lengend Snippet: Induction of NRF2 and NQO1 proteins under oxidative stress is independent of DAP5. Protein extracts of stably transfected NIH-3T3 cells grown in the absence or presence of doxycycline (Dox) for 48 h and untreated or treated with 1 mM H 2 O 2 for 4 h were subjected to western-blotting with the indicated antibodies. The bottom-to-top α–β–γ symbols denote hypo- to hyperphosphorylated 4E-BP1 isoforms.

    Article Snippet: The antibodies used were as follows: anti-eIF4GI and anti-eIF4GII (gifts of Prof. Nahum Sonenberg); anti-DAP5 (CliniSciences #610742); anti-HA-7 (Sigma); anti-β-tubulin (GeneTex #6288022); anti-4E-BP1, anti-NRF2 and anti-p53 (Cell Signaling Technologies #9452, #12721, and #1C12, respectively); anti-Core 20S (Enzo Life Sciences #PW8155); and anti-NQO1 (Santa Cruz #C19).

    Techniques: Stable Transfection, Transfection, Western Blot

    NQO1 Activity of bone marrow cells of guinea pigs fed 0.5 mg or 15 mg vit C/day. (Panel A) AIR, exposed to air; DC, fed 3 mg DC/day; CS, exposed to CS; DC+CS, fed 3 mg DC/day and exposed to CS. * indicates significant difference (p

    Journal: PLoS ONE

    Article Title: NAD(P)H: Quinone Oxidoreductase 1 Deficiency Conjoint with Marginal Vitamin C Deficiency Causes Cigarette Smoke Induced Myelodysplastic Syndromes

    doi: 10.1371/journal.pone.0020590

    Figure Lengend Snippet: NQO1 Activity of bone marrow cells of guinea pigs fed 0.5 mg or 15 mg vit C/day. (Panel A) AIR, exposed to air; DC, fed 3 mg DC/day; CS, exposed to CS; DC+CS, fed 3 mg DC/day and exposed to CS. * indicates significant difference (p

    Article Snippet: Supporting Information NQO1 expression in the bone marrow cells, indicating t hat there is no significant change in NQO1 expression at the protein level. (Panel A) and (Panel B) Bone marrow lysates were separated by 10% SDS PAGE and subjected to immmunobloting using antibody against NQO1.

    Techniques: Activity Assay

    MDS produced in the guinea pigs are irreversible. (Panel A ) Differential staining showing persistent changes in blood and bone marrow cell morphology of MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. A, a–d , represent sham controls (fed 0.5 mg vitamin C/day and exposed to air); A, e–h , represent MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. Blood smear - Leishman stain; bone marrow aspirate - Wright Geimsa stain, except Perls' stain in d and h ; (magnification 400×). (Panel B ) Measurement of CD34(+) cells in bone marrow by flow cytometry. (Panel C ) Geimsa-stained metaphase spread showing aneuploidy in MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day; (magnification 1000×). (Panel D ) NQO1 Activity of bone marrow cells. Bars over the respective columns represent means ± SD (n = 4); * indicates significant difference (p

    Journal: PLoS ONE

    Article Title: NAD(P)H: Quinone Oxidoreductase 1 Deficiency Conjoint with Marginal Vitamin C Deficiency Causes Cigarette Smoke Induced Myelodysplastic Syndromes

    doi: 10.1371/journal.pone.0020590

    Figure Lengend Snippet: MDS produced in the guinea pigs are irreversible. (Panel A ) Differential staining showing persistent changes in blood and bone marrow cell morphology of MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. A, a–d , represent sham controls (fed 0.5 mg vitamin C/day and exposed to air); A, e–h , represent MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. Blood smear - Leishman stain; bone marrow aspirate - Wright Geimsa stain, except Perls' stain in d and h ; (magnification 400×). (Panel B ) Measurement of CD34(+) cells in bone marrow by flow cytometry. (Panel C ) Geimsa-stained metaphase spread showing aneuploidy in MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day; (magnification 1000×). (Panel D ) NQO1 Activity of bone marrow cells. Bars over the respective columns represent means ± SD (n = 4); * indicates significant difference (p

    Article Snippet: Supporting Information NQO1 expression in the bone marrow cells, indicating t hat there is no significant change in NQO1 expression at the protein level. (Panel A) and (Panel B) Bone marrow lysates were separated by 10% SDS PAGE and subjected to immmunobloting using antibody against NQO1.

    Techniques: Produced, Staining, Leishman Stain, Flow Cytometry, Cytometry, Activity Assay

    Immunofluorescent staining of NQO1 protein in BxPc-3 cells. NQO1 protein located in the cytoplasm and nucleus of BxPc-3 cells (green indicates NQO1; blue indicates DAPI; original magnification, ×630). NQO1, nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase 1.

    Journal: Oncology Letters

    Article Title: Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma

    doi: 10.3892/ol.2017.5821

    Figure Lengend Snippet: Immunofluorescent staining of NQO1 protein in BxPc-3 cells. NQO1 protein located in the cytoplasm and nucleus of BxPc-3 cells (green indicates NQO1; blue indicates DAPI; original magnification, ×630). NQO1, nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase 1.

    Article Snippet: The cells were subsequently incubated with anti-NQO1 antibody (dilution, 1:200; cat. no. A180:sc-32793; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight.

    Techniques: Staining

    Immunohistochemical staining of NQO1 protein expression in PDAC tissues. (A) NQO1 protein expression was evaluated in microarray of PDAC tissues. (B) NQO1 staining demonstrated negative expression in non-tumor tissues. (C) Strongly positive expression of NQO1 protein in PDAC with metastasis. (D) NQO1 protein was weakly positive expressed in PDAC. (E) NQO1 protein was negatively expressed in PDAC (Original magnification, ×200 in B-E). NQO1, nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase 1; PDAC, pancreatic ductal adenocarcinoma.

    Journal: Oncology Letters

    Article Title: Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma

    doi: 10.3892/ol.2017.5821

    Figure Lengend Snippet: Immunohistochemical staining of NQO1 protein expression in PDAC tissues. (A) NQO1 protein expression was evaluated in microarray of PDAC tissues. (B) NQO1 staining demonstrated negative expression in non-tumor tissues. (C) Strongly positive expression of NQO1 protein in PDAC with metastasis. (D) NQO1 protein was weakly positive expressed in PDAC. (E) NQO1 protein was negatively expressed in PDAC (Original magnification, ×200 in B-E). NQO1, nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase 1; PDAC, pancreatic ductal adenocarcinoma.

    Article Snippet: The cells were subsequently incubated with anti-NQO1 antibody (dilution, 1:200; cat. no. A180:sc-32793; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight.

    Techniques: Immunohistochemistry, Staining, Expressing, Microarray

    Kaplan-Meier survival curves of PDAC patients in early and late stage. (A) Kaplan-Meier curves for OS in early-stage PDAC patients with low and high level of NQO1 expression (log-rank=18.402, P

    Journal: Oncology Letters

    Article Title: Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma

    doi: 10.3892/ol.2017.5821

    Figure Lengend Snippet: Kaplan-Meier survival curves of PDAC patients in early and late stage. (A) Kaplan-Meier curves for OS in early-stage PDAC patients with low and high level of NQO1 expression (log-rank=18.402, P

    Article Snippet: The cells were subsequently incubated with anti-NQO1 antibody (dilution, 1:200; cat. no. A180:sc-32793; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight.

    Techniques: Expressing

    Kaplan-Meier survival curves illustrating the significance of NQO1 expression in PDAC patients with different grading. (A) OS rates of patients with high (solid; n=83) and low (dashed; n=43) NQO1 expression (P

    Journal: Oncology Letters

    Article Title: Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma

    doi: 10.3892/ol.2017.5821

    Figure Lengend Snippet: Kaplan-Meier survival curves illustrating the significance of NQO1 expression in PDAC patients with different grading. (A) OS rates of patients with high (solid; n=83) and low (dashed; n=43) NQO1 expression (P

    Article Snippet: The cells were subsequently incubated with anti-NQO1 antibody (dilution, 1:200; cat. no. A180:sc-32793; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight.

    Techniques: Expressing

    Association between NQO1 expression and clinicopathological significance of PDAC. The expression level of NQO1 protein was significantly associated with (A) grading (P

    Journal: Oncology Letters

    Article Title: Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma

    doi: 10.3892/ol.2017.5821

    Figure Lengend Snippet: Association between NQO1 expression and clinicopathological significance of PDAC. The expression level of NQO1 protein was significantly associated with (A) grading (P

    Article Snippet: The cells were subsequently incubated with anti-NQO1 antibody (dilution, 1:200; cat. no. A180:sc-32793; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight.

    Techniques: Expressing

    Kaplan-Meier survival curves of PDAC patients with lymph node metastasis and without metastasis. (A) Kaplan-Meier curves for OS in the absence of LN metastasis in PDAC patients with low and high level of NQO1 expression (log-rank=18.402, P

    Journal: Oncology Letters

    Article Title: Clinicopathological implications of NQO1 overexpression in the prognosis of pancreatic adenocarcinoma

    doi: 10.3892/ol.2017.5821

    Figure Lengend Snippet: Kaplan-Meier survival curves of PDAC patients with lymph node metastasis and without metastasis. (A) Kaplan-Meier curves for OS in the absence of LN metastasis in PDAC patients with low and high level of NQO1 expression (log-rank=18.402, P

    Article Snippet: The cells were subsequently incubated with anti-NQO1 antibody (dilution, 1:200; cat. no. A180:sc-32793; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight.

    Techniques: Expressing

    NQO1 inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Stable Transfection, Expressing, Transfection

    NQO1 knockdown in myoblasts reduces steady-state PGC-1α protein levels and activity. (A) NQO1 was knocked down in C2C12 cells using a lentivirus-based approach (3 individual experiments); all were run on the same gel, and cells were analyzed for

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 knockdown in myoblasts reduces steady-state PGC-1α protein levels and activity. (A) NQO1 was knocked down in C2C12 cells using a lentivirus-based approach (3 individual experiments); all were run on the same gel, and cells were analyzed for

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Activity Assay

    NQO1 protects PGC-1α by NADH-dependent interaction. (A) HEK-293 cells were transfected as indicated and harvested for analysis 24 h posttransfection. HA beads were used to immunoprecipitate (IP) HA-tagged PGC-1α, whereas HA-p73β

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 protects PGC-1α by NADH-dependent interaction. (A) HEK-293 cells were transfected as indicated and harvested for analysis 24 h posttransfection. HA beads were used to immunoprecipitate (IP) HA-tagged PGC-1α, whereas HA-p73β

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Transfection

    PGC-1α is an intrinsically disordered protein by prediction, susceptible to in vitro degradation by the 20S PC and protected by NQO1. (A) Analysis of PGC-1α and PCNA amino acid sequences by the FoldIndex prediction program. (B) In vitro

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: PGC-1α is an intrinsically disordered protein by prediction, susceptible to in vitro degradation by the 20S PC and protected by NQO1. (A) Analysis of PGC-1α and PCNA amino acid sequences by the FoldIndex prediction program. (B) In vitro

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, In Vitro

    Schematic model illustrating the convergent actions of CREB, NQO1, AMPK, and Sirt1 on PGC-1α. The scheme summarizes PGC-1α transcription and posttranslational regulation by different known metabolite-sensing proteins, including NQO1.

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: Schematic model illustrating the convergent actions of CREB, NQO1, AMPK, and Sirt1 on PGC-1α. The scheme summarizes PGC-1α transcription and posttranslational regulation by different known metabolite-sensing proteins, including NQO1.

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography

    A role for NQO1 activity in regulating PGC-1α in fasting liver. (A) Mice were either fed ad libitum or fasted during the indicated time points (starting from the beginning of the night phase) and then sacrificed for hepatic analysis of NADH and

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: A role for NQO1 activity in regulating PGC-1α in fasting liver. (A) Mice were either fed ad libitum or fasted during the indicated time points (starting from the beginning of the night phase) and then sacrificed for hepatic analysis of NADH and

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Activity Assay, Pyrolysis Gas Chromatography, Mouse Assay

    NQO1 and PGC-1α steady-state levels in myoblasts and myotubes. (A) Analysis of PGC-1α and NQO1 protein and mRNA levels in C2C12 myoblast (MB) cells and at sequential days after the cells were put in differentiation medium. (B) Same as

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 and PGC-1α steady-state levels in myoblasts and myotubes. (A) Analysis of PGC-1α and NQO1 protein and mRNA levels in C2C12 myoblast (MB) cells and at sequential days after the cells were put in differentiation medium. (B) Same as

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography

    NQO1 regulates PGC-1α accumulation in response to induction by starvation-mimicking conditions in mouse primary hepatocytes. (A) Primary hepatocytes were infected with either GFP- or NQO1-expressing adenoviruses. Expression levels of NQO1 mRNA

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 regulates PGC-1α accumulation in response to induction by starvation-mimicking conditions in mouse primary hepatocytes. (A) Primary hepatocytes were infected with either GFP- or NQO1-expressing adenoviruses. Expression levels of NQO1 mRNA

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Infection, Expressing