Structured Review

Santa Cruz Biotechnology anti ho 1
Digoxin decreased Nrf2 at transcriptional level through inhibiting PI3K/Akt pathway in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on protein levels of p-JNK, JNK, p-ERK1/2, ERK1/2, p-P38 and P38. (C–D) Effects of digoxin on protein levels of PI3K, p-Akt, Akt. (E–F) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2, NQO1, <t>HO-1,</t> and GCLC were detected by Western blot. (G–H) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, Nrf2 mRNA levels were detected by qRT-PCR. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P
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1) Product Images from "Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway"

Article Title: Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway

Journal: Redox Biology

doi: 10.1016/j.redox.2019.101131

Digoxin decreased Nrf2 at transcriptional level through inhibiting PI3K/Akt pathway in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on protein levels of p-JNK, JNK, p-ERK1/2, ERK1/2, p-P38 and P38. (C–D) Effects of digoxin on protein levels of PI3K, p-Akt, Akt. (E–F) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2, NQO1, HO-1, and GCLC were detected by Western blot. (G–H) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, Nrf2 mRNA levels were detected by qRT-PCR. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P
Figure Legend Snippet: Digoxin decreased Nrf2 at transcriptional level through inhibiting PI3K/Akt pathway in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on protein levels of p-JNK, JNK, p-ERK1/2, ERK1/2, p-P38 and P38. (C–D) Effects of digoxin on protein levels of PI3K, p-Akt, Akt. (E–F) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2, NQO1, HO-1, and GCLC were detected by Western blot. (G–H) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, Nrf2 mRNA levels were detected by qRT-PCR. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

Techniques Used: Western Blot, Quantitative RT-PCR

Digoxin increased the sensitivity of SW1990/Gem and Panc-1/Gem cells to gemcitabine by inhibiting Nrf2 signaling. (A–B) Effects of Nrf2 knockdown on protein levels of Nrf2, NQO1, HO-1, and GCLC in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of Nrf2 knockdown on reversing drug resistance of gemcitabine in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P
Figure Legend Snippet: Digoxin increased the sensitivity of SW1990/Gem and Panc-1/Gem cells to gemcitabine by inhibiting Nrf2 signaling. (A–B) Effects of Nrf2 knockdown on protein levels of Nrf2, NQO1, HO-1, and GCLC in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of Nrf2 knockdown on reversing drug resistance of gemcitabine in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

Techniques Used:

Nrf2 signaling was upregulated in SW1990/Gem and Panc-1/Gem cells. (A–B) The cytotoxicity of gemcitabine to SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (C–D) Western blot was used to detect the protein level of Nrf2, NQO1, HO-1, and GCLC in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (E–F) The expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P
Figure Legend Snippet: Nrf2 signaling was upregulated in SW1990/Gem and Panc-1/Gem cells. (A–B) The cytotoxicity of gemcitabine to SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (C–D) Western blot was used to detect the protein level of Nrf2, NQO1, HO-1, and GCLC in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (E–F) The expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

Techniques Used: Western Blot, Expressing

Digoxin inhibited the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of digoxin on protein levels of NQO1, HO-1 and GCLC in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as **P
Figure Legend Snippet: Digoxin inhibited the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of digoxin on protein levels of NQO1, HO-1 and GCLC in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as **P

Techniques Used: Expressing

Digoxin sensitized SW1990/Gem cells-derived xenografts to gemcitabine treatment by inhibiting Nrf2 signaling. (A–D) Digoxin sensitized SW1990/Gem-shControl cells-derived xenografts to gemcitabine treatment. (E–H) Digoxin could not sensitize SW1990/Gem-shNrf2 cells-derived xenografts to gemcitabine treatment. (I) Effects of digoxin on the protein levels of Nrf2, NQO1, HO-1, and GCLC in tumor tissues. (J) Tumor tissues were subjected to IHC-Nrf2, IHC-NQO1, IHC-HO-1, IHC-GCLC, IHC-Ki67 and TUNEL staining. All images were shown at ×200. Data were expressed as mean ± SD, n = 6. Significant differences were indicated as ***P
Figure Legend Snippet: Digoxin sensitized SW1990/Gem cells-derived xenografts to gemcitabine treatment by inhibiting Nrf2 signaling. (A–D) Digoxin sensitized SW1990/Gem-shControl cells-derived xenografts to gemcitabine treatment. (E–H) Digoxin could not sensitize SW1990/Gem-shNrf2 cells-derived xenografts to gemcitabine treatment. (I) Effects of digoxin on the protein levels of Nrf2, NQO1, HO-1, and GCLC in tumor tissues. (J) Tumor tissues were subjected to IHC-Nrf2, IHC-NQO1, IHC-HO-1, IHC-GCLC, IHC-Ki67 and TUNEL staining. All images were shown at ×200. Data were expressed as mean ± SD, n = 6. Significant differences were indicated as ***P

Techniques Used: Derivative Assay, Immunohistochemistry, TUNEL Assay, Staining

2) Product Images from "Identification and Functional Studies of a New Nrf2 Partner IQGAP1: A Critical Role in the Stability and Transactivation of Nrf2"

Article Title: Identification and Functional Studies of a New Nrf2 Partner IQGAP1: A Critical Role in the Stability and Transactivation of Nrf2

Journal: Antioxidants & Redox Signaling

doi: 10.1089/ars.2012.4586

Translocation of Nrf2-IQGAP1 complex into the nucleus by calcium treatment and induction of HO-1 protein. (A) To verify the involvement of calcium in endogenous Nrf2-IQGAP1 complexes, HeLa cells were transfected with pEGFP-Nrf2 (8 μg/100 mm
Figure Legend Snippet: Translocation of Nrf2-IQGAP1 complex into the nucleus by calcium treatment and induction of HO-1 protein. (A) To verify the involvement of calcium in endogenous Nrf2-IQGAP1 complexes, HeLa cells were transfected with pEGFP-Nrf2 (8 μg/100 mm

Techniques Used: Translocation Assay, Transfection

IQGAP1 enhances the expression of Nrf2 and induction of HO-1 expression as well as the stability of Nrf2. (A) Based on mass data shown in , the newly identified IQGAP1 was then tested to see whether it could regulate the expression of HO-1, which
Figure Legend Snippet: IQGAP1 enhances the expression of Nrf2 and induction of HO-1 expression as well as the stability of Nrf2. (A) Based on mass data shown in , the newly identified IQGAP1 was then tested to see whether it could regulate the expression of HO-1, which

Techniques Used: Expressing

siIQGAP1 decreases the stability of Nrf2 and attenuates HO-1 expression and ARE-luciferase activity. (A) HeLa cells were transfected with siIQGAP1 for 72 h using Lipofectamin 2000 reagent to silence or knockdown the endogenous IQGAP1 gene expression.
Figure Legend Snippet: siIQGAP1 decreases the stability of Nrf2 and attenuates HO-1 expression and ARE-luciferase activity. (A) HeLa cells were transfected with siIQGAP1 for 72 h using Lipofectamin 2000 reagent to silence or knockdown the endogenous IQGAP1 gene expression.

Techniques Used: Expressing, Luciferase, Activity Assay, Transfection

3) Product Images from "Ethyl Pyruvate Induces Heme Oxygenase-1 Through p38 Mitogen-Activated Protein Kinase Activation by Depletion of Glutathione in RAW 264.7 Cells and Improves Survival in Septic Animals"

Article Title: Ethyl Pyruvate Induces Heme Oxygenase-1 Through p38 Mitogen-Activated Protein Kinase Activation by Depletion of Glutathione in RAW 264.7 Cells and Improves Survival in Septic Animals

Journal: Antioxidants & Redox Signaling

doi: 10.1089/ars.2011.3994

Importance of HO-1 in the anti-inflammatory action of EP. Cells were transfected with ssiRNA or siHO-1 RNA, which were subjected to western blot to confirm siHO-1 efficiency by EP (25 m M ) for 8 h (A) . Cells were stimulated with LPS (1 μg/ml)
Figure Legend Snippet: Importance of HO-1 in the anti-inflammatory action of EP. Cells were transfected with ssiRNA or siHO-1 RNA, which were subjected to western blot to confirm siHO-1 efficiency by EP (25 m M ) for 8 h (A) . Cells were stimulated with LPS (1 μg/ml)

Techniques Used: Transfection, Western Blot

Involvement of Nrf2 in EP-mediated HO-1 expression. Cells were treated for 1 h with EP at 5, 10, and 25 m M , and then harvested and subjected to cytosol/nuclear fractionation (A) . Cells were transiently transfected with the ARE-luc vector
Figure Legend Snippet: Involvement of Nrf2 in EP-mediated HO-1 expression. Cells were treated for 1 h with EP at 5, 10, and 25 m M , and then harvested and subjected to cytosol/nuclear fractionation (A) . Cells were transiently transfected with the ARE-luc vector

Techniques Used: Expressing, Fractionation, Transfection, Plasmid Preparation

Effect of EP on the expression of HO-1 in macrophages. Cells were treated with EP at doses 5, 10, and 25 m M for 8 h (A) or at dose 25 m M for 1, 2, 4, 6, 8, 12, and 24 h (B) . After incubation, cells were harvested and subjected
Figure Legend Snippet: Effect of EP on the expression of HO-1 in macrophages. Cells were treated with EP at doses 5, 10, and 25 m M for 8 h (A) or at dose 25 m M for 1, 2, 4, 6, 8, 12, and 24 h (B) . After incubation, cells were harvested and subjected

Techniques Used: Expressing, Incubation

EP fails to increase the survival rate in the CLP-induced sepsis model of HO-1 −/− mice. BALB/c WT ( n =12) and HO-1 −/− mice ( n =12) were subjected to CLP with EP ( n =6, 40 mg/kg i.p.) or an equal volume of vehicle (
Figure Legend Snippet: EP fails to increase the survival rate in the CLP-induced sepsis model of HO-1 −/− mice. BALB/c WT ( n =12) and HO-1 −/− mice ( n =12) were subjected to CLP with EP ( n =6, 40 mg/kg i.p.) or an equal volume of vehicle (

Techniques Used: Mouse Assay

EP induces HO-1 through p38 MAPK. Cells were pretreated with SB203580 (10 μ M ), SP600125 (10 μ M ), and PD98059 (10 μ M ) for 1 h, and then cells were treated with EP (25 m M ) for 8 h
Figure Legend Snippet: EP induces HO-1 through p38 MAPK. Cells were pretreated with SB203580 (10 μ M ), SP600125 (10 μ M ), and PD98059 (10 μ M ) for 1 h, and then cells were treated with EP (25 m M ) for 8 h

Techniques Used:

4) Product Images from "Association of HMGB1 with oxidative stress markers and regulators in PDR"

Article Title: Association of HMGB1 with oxidative stress markers and regulators in PDR

Journal: Molecular Vision

doi:

Effects of HMGB1 on the expressions of VAP-1, 8-OHdG, and HO-1 in human retinal microvascular endothelial cells. Immunofluorescence microscopy showed that HMGB1 treatment induced membranous VAP-1 translocation ( A ), the upregulation of cytoplasmic and nuclear immunoreactivities for 8-OHdG ( B ), and cytoplasmic immunoreactivity for HO-1 ( C ; green). Nuclei were counterstained with DAPI (blue).
Figure Legend Snippet: Effects of HMGB1 on the expressions of VAP-1, 8-OHdG, and HO-1 in human retinal microvascular endothelial cells. Immunofluorescence microscopy showed that HMGB1 treatment induced membranous VAP-1 translocation ( A ), the upregulation of cytoplasmic and nuclear immunoreactivities for 8-OHdG ( B ), and cytoplasmic immunoreactivity for HO-1 ( C ; green). Nuclei were counterstained with DAPI (blue).

Techniques Used: Immunofluorescence, Microscopy, Translocation Assay

5) Product Images from "Protective effect of higenamine ameliorates collagen-induced arthritis through heme oxygenase-1 and PI3K/Akt/Nrf-2 signaling pathways"

Article Title: Protective effect of higenamine ameliorates collagen-induced arthritis through heme oxygenase-1 and PI3K/Akt/Nrf-2 signaling pathways

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2016.3730

Protective effect of HIG on the HO-1 protein expression by (A) western blotting assays and (B) statistical analysis of HO-1 protein expression. ## P
Figure Legend Snippet: Protective effect of HIG on the HO-1 protein expression by (A) western blotting assays and (B) statistical analysis of HO-1 protein expression. ## P

Techniques Used: Expressing, Western Blot

6) Product Images from "Hydrogen Sulfide Alleviates Diabetic Nephropathy in a Streptozotocin-induced Diabetic Rat Model"

Article Title: Hydrogen Sulfide Alleviates Diabetic Nephropathy in a Streptozotocin-induced Diabetic Rat Model

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M114.596593

Representative immunoblots and densitometric analysis of Nrf2 in the nucleus and cytosol ( A ) and its downstream targets HO-1 and NQO1 ( B ). *, p
Figure Legend Snippet: Representative immunoblots and densitometric analysis of Nrf2 in the nucleus and cytosol ( A ) and its downstream targets HO-1 and NQO1 ( B ). *, p

Techniques Used: Western Blot

7) Product Images from "Ursolic acid sensitizes cisplatin-resistant HepG2/DDP cells to cisplatin via inhibiting Nrf2/ARE pathway"

Article Title: Ursolic acid sensitizes cisplatin-resistant HepG2/DDP cells to cisplatin via inhibiting Nrf2/ARE pathway

Journal: Drug Design, Development and Therapy

doi: 10.2147/DDDT.S110505

UA sensitizes HepG2/DDP cells to low-dose cisplatin via inhibition of Nrf2/ARE signaling pathway. Notes: ( A ) HepG2/DDP cells were transfected with Nrf2 siRNA (si-Nrf2) or negative control (si-Con), or ( C ) HepG2/DDP cells were transfected with Nrf2 cDNA or empty vector (Vector), then treated with 8.92 μg/mL cisplatin (IC 30 of cisplatin for HepG2/DDP cells) and/or UA (2.25 μg/mL) for 48 hours. The level of Nrf2, HO-1, NQO1, and GST was detected by Western blot analysis. ( B ) HepG2/DDP cells were transfected with si-Nrf2 or si-Con, or ( D ) HepG2/DDP cells were transfected with Nrf2 cDNA or empty vector (Vector), then treated with series concentration of cisplatin (2–512 μg/mL) and/or UA (2.25 μg/mL) for 48 hours. The inhibition rate of cell was detected by CCK8 assay. Results are representative of three different experiments, and they are expressed as mean ± SD. Abbreviations: ARE, antioxidant response element; CCK8, Cell Counting Kit 8; cDNA, complementary DNA; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GST, glutathione S -transferase; HepG2/DDP, cisplatin–resistant hepatocellular carcinoma cell line; HO-1, heme oxygenase-1; IC 30 , 30% inhibitory concentration; NQO1, NAD(P)H quinone oxidoreductase 1; Nrf2, nuclear factor erythroid-2-related factor 2; SD, standard deviation; siRNA, small interfering RNA; UA, ursolic acid.
Figure Legend Snippet: UA sensitizes HepG2/DDP cells to low-dose cisplatin via inhibition of Nrf2/ARE signaling pathway. Notes: ( A ) HepG2/DDP cells were transfected with Nrf2 siRNA (si-Nrf2) or negative control (si-Con), or ( C ) HepG2/DDP cells were transfected with Nrf2 cDNA or empty vector (Vector), then treated with 8.92 μg/mL cisplatin (IC 30 of cisplatin for HepG2/DDP cells) and/or UA (2.25 μg/mL) for 48 hours. The level of Nrf2, HO-1, NQO1, and GST was detected by Western blot analysis. ( B ) HepG2/DDP cells were transfected with si-Nrf2 or si-Con, or ( D ) HepG2/DDP cells were transfected with Nrf2 cDNA or empty vector (Vector), then treated with series concentration of cisplatin (2–512 μg/mL) and/or UA (2.25 μg/mL) for 48 hours. The inhibition rate of cell was detected by CCK8 assay. Results are representative of three different experiments, and they are expressed as mean ± SD. Abbreviations: ARE, antioxidant response element; CCK8, Cell Counting Kit 8; cDNA, complementary DNA; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GST, glutathione S -transferase; HepG2/DDP, cisplatin–resistant hepatocellular carcinoma cell line; HO-1, heme oxygenase-1; IC 30 , 30% inhibitory concentration; NQO1, NAD(P)H quinone oxidoreductase 1; Nrf2, nuclear factor erythroid-2-related factor 2; SD, standard deviation; siRNA, small interfering RNA; UA, ursolic acid.

Techniques Used: Inhibition, Transfection, Negative Control, Plasmid Preparation, Western Blot, Concentration Assay, CCK-8 Assay, Cell Counting, Standard Deviation, Small Interfering RNA

Nrf2 was overexpressed in cisplatin-resistant human hepatocellular carcinoma HepG2/DDP cells. Notes: ( A ) HepG2 cells were treated with series concentration of cisplatin (0.1–25.6 μg/mL) for 48 hours. ( B ) HepG2/DDP cells were treated with series concentration of cisplatin (2–512 μg/mL) for 48 hours. ( C ) The level of Nrf2 and its downstream target genes HO-1, NQO1, and GST in HepG2 and HepG2/DDP cells was detected by Western blot assay. Results are representative of three different experiments, and they are expressed as mean ± SD. Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GST, glutathione S -transferase; HepG2, hepatocellular carcinoma cell line; HepG2/DDP, cisplatin-resistant hepatocellular carcinoma cell line; HO-1, heme oxygenase-1; NQO1, NAD(P)H quinone oxidoreductase 1; Nrf2, nuclear factor erythroid-2-related factor 2; SD, standard deviation.
Figure Legend Snippet: Nrf2 was overexpressed in cisplatin-resistant human hepatocellular carcinoma HepG2/DDP cells. Notes: ( A ) HepG2 cells were treated with series concentration of cisplatin (0.1–25.6 μg/mL) for 48 hours. ( B ) HepG2/DDP cells were treated with series concentration of cisplatin (2–512 μg/mL) for 48 hours. ( C ) The level of Nrf2 and its downstream target genes HO-1, NQO1, and GST in HepG2 and HepG2/DDP cells was detected by Western blot assay. Results are representative of three different experiments, and they are expressed as mean ± SD. Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GST, glutathione S -transferase; HepG2, hepatocellular carcinoma cell line; HepG2/DDP, cisplatin-resistant hepatocellular carcinoma cell line; HO-1, heme oxygenase-1; NQO1, NAD(P)H quinone oxidoreductase 1; Nrf2, nuclear factor erythroid-2-related factor 2; SD, standard deviation.

Techniques Used: Concentration Assay, Western Blot, Standard Deviation

UA–cisplatin combination downregulates Nrf2 and its substrates. Notes: The protein expression levels of Nrf2, HO-1, NQO1, and GST of HepG2/DDP cells treated with 8.92 μg/mL cisplatin and/or UA (2.25 μg/mL) for 48 hours were detected by Western blot analysis. Results are representative of three different experiments, and they are expressed as mean ± SD. Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GST, glutathione S -transferase; HepG2/DDP, cisplatin-resistant hepatocellular carcinoma cell line; HO-1, heme oxygenase-1; NQO1, NAD(P)H quinone oxidoreductase 1; Nrf2, nuclear factor erythroid-2-related factor 2; SD, standard deviation; UA, ursolic acid.
Figure Legend Snippet: UA–cisplatin combination downregulates Nrf2 and its substrates. Notes: The protein expression levels of Nrf2, HO-1, NQO1, and GST of HepG2/DDP cells treated with 8.92 μg/mL cisplatin and/or UA (2.25 μg/mL) for 48 hours were detected by Western blot analysis. Results are representative of three different experiments, and they are expressed as mean ± SD. Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GST, glutathione S -transferase; HepG2/DDP, cisplatin-resistant hepatocellular carcinoma cell line; HO-1, heme oxygenase-1; NQO1, NAD(P)H quinone oxidoreductase 1; Nrf2, nuclear factor erythroid-2-related factor 2; SD, standard deviation; UA, ursolic acid.

Techniques Used: Expressing, Western Blot, Standard Deviation

8) Product Images from "Reciprocal regulation of eNOS, H2S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin"

Article Title: Reciprocal regulation of eNOS, H2S and CO-synthesizing enzymes in human atheroma: Correlation with plaque stability and effects of simvastatin

Journal: Redox Biology

doi: 10.1016/j.redox.2017.02.006

Simvastatin treatment reduces HIF-1α expression and its downstream targets HO-1 and iNOS promoting atheroma stability. Representative western blots and relative densitometric analysis of Α. HIF-1α/β-tubulin B. NOX-4/β-actin C. Immunohistochemical (IHC) analysis of NOX-4 in serial sections from s/nost (n=8) and s/st (n=8) groups D. HO-1/β-actin E. iNOS/β-actin F. Immunohistochemical (IHC) analysis of iNOS in serial sections from s/nost (n=8) and s/st (n=8) groups. For IHC purposes, frames in 200x magnification pictures represent the areas of 400x magnification.
Figure Legend Snippet: Simvastatin treatment reduces HIF-1α expression and its downstream targets HO-1 and iNOS promoting atheroma stability. Representative western blots and relative densitometric analysis of Α. HIF-1α/β-tubulin B. NOX-4/β-actin C. Immunohistochemical (IHC) analysis of NOX-4 in serial sections from s/nost (n=8) and s/st (n=8) groups D. HO-1/β-actin E. iNOS/β-actin F. Immunohistochemical (IHC) analysis of iNOS in serial sections from s/nost (n=8) and s/st (n=8) groups. For IHC purposes, frames in 200x magnification pictures represent the areas of 400x magnification.

Techniques Used: Expressing, Western Blot, Immunohistochemistry

Changes in HIF-1α and its downstream targets NOX-4, HO-1 and iNOS in human atherosclerotic plaques. Representative western blots and relative densitometric analysis of Α. HIF-1α/β-tubulin B. NOX-4/β-actin C. Immunohistochemical (IHC) analysis of NOX-4 in serial sections from stable (n=8) and unstable plaques (n=8) D. HO-1/β-actin E. iNOS/β-actin F. Immunohistochemical (IHC) analysis of iNOS in serial sections from stable (n=8) and unstable plaques (n=8) (*p
Figure Legend Snippet: Changes in HIF-1α and its downstream targets NOX-4, HO-1 and iNOS in human atherosclerotic plaques. Representative western blots and relative densitometric analysis of Α. HIF-1α/β-tubulin B. NOX-4/β-actin C. Immunohistochemical (IHC) analysis of NOX-4 in serial sections from stable (n=8) and unstable plaques (n=8) D. HO-1/β-actin E. iNOS/β-actin F. Immunohistochemical (IHC) analysis of iNOS in serial sections from stable (n=8) and unstable plaques (n=8) (*p

Techniques Used: Western Blot, Immunohistochemistry

Reciprocal regulation of eNOS vs H 2 S in atherosclerotic plaques. eNOS-derived NO increases bioavailability of NO in stable plaques, which would be expected to occur due to eNOS upregulation and phosphorylation (due to Akt activation); concomitantly iNOS and NOX-4 are downregulated presumably due to reduced HIF-1α. Moreover CBS and CSE positively correlate with plaque vulnerability. Simvastatin administration decreases HIF-1α expression and its downstream targets, iNOS and HO-1, whereas it increases the phosphorylation and expression of eNOS. Reduced CSE expression is associated with simvastatin administration and enhanced plaque stability (Modified by [69] ). Red color indicates signaling molecules that decreased, whereas green color indicates signaling molecules that are upregulated.
Figure Legend Snippet: Reciprocal regulation of eNOS vs H 2 S in atherosclerotic plaques. eNOS-derived NO increases bioavailability of NO in stable plaques, which would be expected to occur due to eNOS upregulation and phosphorylation (due to Akt activation); concomitantly iNOS and NOX-4 are downregulated presumably due to reduced HIF-1α. Moreover CBS and CSE positively correlate with plaque vulnerability. Simvastatin administration decreases HIF-1α expression and its downstream targets, iNOS and HO-1, whereas it increases the phosphorylation and expression of eNOS. Reduced CSE expression is associated with simvastatin administration and enhanced plaque stability (Modified by [69] ). Red color indicates signaling molecules that decreased, whereas green color indicates signaling molecules that are upregulated.

Techniques Used: Derivative Assay, Activation Assay, Expressing, Modification

9) Product Images from "Taxifolin Activates the Nrf2 Anti-Oxidative Stress Pathway in Mouse Skin Epidermal JB6 P+ Cells through Epigenetic Modifications"

Article Title: Taxifolin Activates the Nrf2 Anti-Oxidative Stress Pathway in Mouse Skin Epidermal JB6 P+ Cells through Epigenetic Modifications

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms18071546

Effect of TAX on mRNA and protein expression of Nrf2 and Nrf2 target genes (HO-1 and NQO1) in JB6 P+ cells. JB6 P+ cells were treated with different TAX concentration (10 to 40 μM). ( A ) Western blot images of Nrf2 and its downstream genes including HO-1 and NQO1; ( B ) TAX increased the protein expression of Nrf2 and Nrf2 downstream enzymes; ( C ) TAX increased the mRNA levels of Nrf2 and its downstream representative enzymes including HO-1 and NQO1. The graphical data are presented as the mean ± SD from three independent experiments. * p
Figure Legend Snippet: Effect of TAX on mRNA and protein expression of Nrf2 and Nrf2 target genes (HO-1 and NQO1) in JB6 P+ cells. JB6 P+ cells were treated with different TAX concentration (10 to 40 μM). ( A ) Western blot images of Nrf2 and its downstream genes including HO-1 and NQO1; ( B ) TAX increased the protein expression of Nrf2 and Nrf2 downstream enzymes; ( C ) TAX increased the mRNA levels of Nrf2 and its downstream representative enzymes including HO-1 and NQO1. The graphical data are presented as the mean ± SD from three independent experiments. * p

Techniques Used: Expressing, Concentration Assay, Western Blot

10) Product Images from "Anti-Inflammatory and Antioxidant Properties of Dehydrated Potato-Derived Bioactive Compounds in Intestinal Cells"

Article Title: Anti-Inflammatory and Antioxidant Properties of Dehydrated Potato-Derived Bioactive Compounds in Intestinal Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms20236087

Effect of the three fractions (1–10 µg/mL) on HO-1 ( a ) and SOD ( b ) expression in LPS + IFN-stimulated IEC-6 cells. The three tested fractions significantly increased HO-1 ( p
Figure Legend Snippet: Effect of the three fractions (1–10 µg/mL) on HO-1 ( a ) and SOD ( b ) expression in LPS + IFN-stimulated IEC-6 cells. The three tested fractions significantly increased HO-1 ( p

Techniques Used: Expressing

11) Product Images from "Mechanotransduction Drives Post Ischemic Revascularization Through KATP Channel Closure and Production of Reactive Oxygen Species"

Article Title: Mechanotransduction Drives Post Ischemic Revascularization Through KATP Channel Closure and Production of Reactive Oxygen Species

Journal: Antioxidants & Redox Signaling

doi: 10.1089/ars.2012.4971

HO-1 in ischemic tissue in vivo and ischemic cells in vitro. (A) Hind limb muscle tissue and vascular tissue from the site of ligation to the knee was excised 1 (Isc, 1d) and 5 days (Isc, 5d) post ligation. The tissue was fixed, sectioned, and immunostained
Figure Legend Snippet: HO-1 in ischemic tissue in vivo and ischemic cells in vitro. (A) Hind limb muscle tissue and vascular tissue from the site of ligation to the knee was excised 1 (Isc, 1d) and 5 days (Isc, 5d) post ligation. The tissue was fixed, sectioned, and immunostained

Techniques Used: In Vivo, In Vitro, Ligation

12) Product Images from "Brain deposition and neurotoxicity of manganese in adult mice exposed via the drinking water"

Article Title: Brain deposition and neurotoxicity of manganese in adult mice exposed via the drinking water

Journal: Archives of toxicology

doi: 10.1007/s00204-013-1088-3

Nigral mRNA expression Effect of Mn after 8 weeks of DW (0.4 g/l) exposure on mRNA expression levels of tyrosine hydroxylase (TH; a), dopamine receptor -2 (D2DR; b), glutamate decarboxylase 1 (GAD1; c), heme oxygenase-1 (HO-1; d), inducible nitric oxide synthase (NOS2; e) and glial fibrillary acidic protein (GFAP; f) in the substantia nigra. mRNA data are β-actin-normalized and are presented as fold change relative to control (mean ± SEM). * Indicates a significant effect of Mn (p ≤ 0.05).
Figure Legend Snippet: Nigral mRNA expression Effect of Mn after 8 weeks of DW (0.4 g/l) exposure on mRNA expression levels of tyrosine hydroxylase (TH; a), dopamine receptor -2 (D2DR; b), glutamate decarboxylase 1 (GAD1; c), heme oxygenase-1 (HO-1; d), inducible nitric oxide synthase (NOS2; e) and glial fibrillary acidic protein (GFAP; f) in the substantia nigra. mRNA data are β-actin-normalized and are presented as fold change relative to control (mean ± SEM). * Indicates a significant effect of Mn (p ≤ 0.05).

Techniques Used: Expressing

13) Product Images from "Interplay between VEGF and Nrf2 regulates angiogenesis due to intracranial venous hypertension"

Article Title: Interplay between VEGF and Nrf2 regulates angiogenesis due to intracranial venous hypertension

Journal: Scientific Reports

doi: 10.1038/srep37338

Nrf2 activator t-BHQ activates VEGF via Nrf2/HO-1/HIF-1α pathways. ( A,B ) After the BMECs was incubated with 1–10 μM of t-BHQ for 6 h, the mRNA and protein levels of Nrf2 and VEGF was determined using β-actin as the loading control by Western blot and qRT-PCR. The histograms show the ratio of Nrf2/β-actin and VEGF/β-actin. ( C ) Western blot shows the upregulation of Nrf2 downstream protein HO-1, NQO1 and HIF-1α in response to t-BHQ. ( D,E ) 10 μM t-BHQ was used to stimulate the BMECs at different time points, and the protein and mRNA levels of VEGF and Nrf2 were measured. ( F ) The expression of HO-1, NQO1, and HIF-1α were also measured and calculated. ( E ) After 48 h of transfection with siRNA–HO-1 or control siRNA, the BMECs were incubated with 10 μM t-BHQ for 6 h. Then, Western blot was used to analyse the expression of HO-1, VEGF and HIF-1α. *p
Figure Legend Snippet: Nrf2 activator t-BHQ activates VEGF via Nrf2/HO-1/HIF-1α pathways. ( A,B ) After the BMECs was incubated with 1–10 μM of t-BHQ for 6 h, the mRNA and protein levels of Nrf2 and VEGF was determined using β-actin as the loading control by Western blot and qRT-PCR. The histograms show the ratio of Nrf2/β-actin and VEGF/β-actin. ( C ) Western blot shows the upregulation of Nrf2 downstream protein HO-1, NQO1 and HIF-1α in response to t-BHQ. ( D,E ) 10 μM t-BHQ was used to stimulate the BMECs at different time points, and the protein and mRNA levels of VEGF and Nrf2 were measured. ( F ) The expression of HO-1, NQO1, and HIF-1α were also measured and calculated. ( E ) After 48 h of transfection with siRNA–HO-1 or control siRNA, the BMECs were incubated with 10 μM t-BHQ for 6 h. Then, Western blot was used to analyse the expression of HO-1, VEGF and HIF-1α. *p

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

The proposed signaling pathway of this study: VH may participate in angiogenesis via activating Nrf2-ARE and HIF-1α/VEGF signaling pathway in AVMs. In vitro , we observed that there existed a VEGF-Nrf2 positive feed back loop in mice BMECs. VEGF activated Nrf2-ARE system via ERK1/2 signaling pathway, in turn, upregulated the expression of itself. Besides, Nrf2 palys an crucial role in the process of angiogenesis and the activator of Nrf2 t-BHQ may also upregulate VEGF expression via Nrf2/HO-1/HIF-1α pathways. Furthermore, knockout of Nrf2 impairs VEGF-induced BMECs migration and angiogenesis ex vivo .
Figure Legend Snippet: The proposed signaling pathway of this study: VH may participate in angiogenesis via activating Nrf2-ARE and HIF-1α/VEGF signaling pathway in AVMs. In vitro , we observed that there existed a VEGF-Nrf2 positive feed back loop in mice BMECs. VEGF activated Nrf2-ARE system via ERK1/2 signaling pathway, in turn, upregulated the expression of itself. Besides, Nrf2 palys an crucial role in the process of angiogenesis and the activator of Nrf2 t-BHQ may also upregulate VEGF expression via Nrf2/HO-1/HIF-1α pathways. Furthermore, knockout of Nrf2 impairs VEGF-induced BMECs migration and angiogenesis ex vivo .

Techniques Used: In Vitro, Mouse Assay, Expressing, Knock-Out, Migration, Ex Vivo

14) Product Images from "AVE0991, a Nonpeptide Compound, Attenuates Angiotensin II-Induced Vascular Smooth Muscle Cell Proliferation via Induction of Heme Oxygenase-1 and Downregulation of p-38 MAPK Phosphorylation"

Article Title: AVE0991, a Nonpeptide Compound, Attenuates Angiotensin II-Induced Vascular Smooth Muscle Cell Proliferation via Induction of Heme Oxygenase-1 and Downregulation of p-38 MAPK Phosphorylation

Journal: International Journal of Hypertension

doi: 10.1155/2012/958298

The effect of AVE0991 on the Ang II-induced VSMCs [ 3 H]thymidine incorporation efficiency with pretreatment with the HO-1 inhibitor ZnPPIX. ** P
Figure Legend Snippet: The effect of AVE0991 on the Ang II-induced VSMCs [ 3 H]thymidine incorporation efficiency with pretreatment with the HO-1 inhibitor ZnPPIX. ** P

Techniques Used:

The effect of AVE0991 on the p38 phosphorylation level in VSMCs when combined with the HO-1 inhibitor ZnPPIX. ** P
Figure Legend Snippet: The effect of AVE0991 on the p38 phosphorylation level in VSMCs when combined with the HO-1 inhibitor ZnPPIX. ** P

Techniques Used:

The effect of AVE0991 on HO-1 protein expression in VSMCs that were induced by Ang II. * P
Figure Legend Snippet: The effect of AVE0991 on HO-1 protein expression in VSMCs that were induced by Ang II. * P

Techniques Used: Expressing

The effect of AVE0991 on Ang II-stimulated ROS production in VSMCs that were pretreated with the HO-1 inhibitor ZnPPIX. * P
Figure Legend Snippet: The effect of AVE0991 on Ang II-stimulated ROS production in VSMCs that were pretreated with the HO-1 inhibitor ZnPPIX. * P

Techniques Used:

15) Product Images from "Anthocyanins encapsulated by PLGA@PEG nanoparticles potentially improved its free radical scavenging capabilities via p38/JNK pathway against Aβ1–42-induced oxidative stress"

Article Title: Anthocyanins encapsulated by PLGA@PEG nanoparticles potentially improved its free radical scavenging capabilities via p38/JNK pathway against Aβ1–42-induced oxidative stress

Journal: Journal of Nanobiotechnology

doi: 10.1186/s12951-016-0227-4

Schematic representation of intracellular uptake of anthocyanin loaded PEGylated NPs by SH-SY5Y cell via endocytosis; inside the cellular cytoplasm, the endosome is broken down by lysosomal enzymes and the drug is released in the cytoplasm and reverts the Aβ 1–42 -induced Aβ pathology by abrogating ROS generation via the P38-MAPK/JNK pathways accompanied by induction of endogenous nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1)
Figure Legend Snippet: Schematic representation of intracellular uptake of anthocyanin loaded PEGylated NPs by SH-SY5Y cell via endocytosis; inside the cellular cytoplasm, the endosome is broken down by lysosomal enzymes and the drug is released in the cytoplasm and reverts the Aβ 1–42 -induced Aβ pathology by abrogating ROS generation via the P38-MAPK/JNK pathways accompanied by induction of endogenous nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1)

Techniques Used:

An-NPs treatment alleviated oxidative stress and upregulated Nrf-2 and its downstream targets genes HO-1 expressions. a Shown are representative Western Blots probed with antibodies of Nrf2, and HO-1 in the Aβ 1–42 treated SHSY-5Y cells. Data are the representative of three individual experiments (n = 3). The protein bands were quantified using sigma gel software. β-Actin was used to show equivalent amounts of protein loading. b The double immunofluorescence images of SH-SY5Y cells after Aβ 1-42 and An-NPs treatment, showing 8-Oxo-G ( green ) and Nrf2 ( red ), proteins and their respective relative density histograms. The DAPI ( blue ) was used to stain the nucleus. All the experiments were performed in triplicate. The details are given in the “ Methods ” section. *Significantly different from the control; # significantly different from Aβ 1–42 -treated group. Significance = **p
Figure Legend Snippet: An-NPs treatment alleviated oxidative stress and upregulated Nrf-2 and its downstream targets genes HO-1 expressions. a Shown are representative Western Blots probed with antibodies of Nrf2, and HO-1 in the Aβ 1–42 treated SHSY-5Y cells. Data are the representative of three individual experiments (n = 3). The protein bands were quantified using sigma gel software. β-Actin was used to show equivalent amounts of protein loading. b The double immunofluorescence images of SH-SY5Y cells after Aβ 1-42 and An-NPs treatment, showing 8-Oxo-G ( green ) and Nrf2 ( red ), proteins and their respective relative density histograms. The DAPI ( blue ) was used to stain the nucleus. All the experiments were performed in triplicate. The details are given in the “ Methods ” section. *Significantly different from the control; # significantly different from Aβ 1–42 -treated group. Significance = **p

Techniques Used: Western Blot, Software, Immunofluorescence, Staining

16) Product Images from "HO-1 Induction by Selaginella tamariscina Extract Inhibits Inflammatory Response in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages"

Article Title: HO-1 Induction by Selaginella tamariscina Extract Inhibits Inflammatory Response in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2018/7816923

Induction of Nrf2/HO-1 signaling pathway by STE. (a) Immunofluorescence images of the nuclear translocation of Nrf2 induced by STE. RAW 264.7 cells were treated with STE (300 μ g/mL) for 3 h. (b) Induction of HO-1 by STE. RAW 264.7 cells were treated with various concentrations of STE (10, 100, or 300 μ g/mL) for 18 h, lysed, and western blotted. (Significant vs. vehicle-treated control, ∗ p
Figure Legend Snippet: Induction of Nrf2/HO-1 signaling pathway by STE. (a) Immunofluorescence images of the nuclear translocation of Nrf2 induced by STE. RAW 264.7 cells were treated with STE (300 μ g/mL) for 3 h. (b) Induction of HO-1 by STE. RAW 264.7 cells were treated with various concentrations of STE (10, 100, or 300 μ g/mL) for 18 h, lysed, and western blotted. (Significant vs. vehicle-treated control, ∗ p

Techniques Used: Immunofluorescence, Translocation Assay, Western Blot

17) Product Images from "HO-1 Induction by CO-RM2 Attenuates TNF-α-Induced Cytosolic Phospholipase A2 Expression via Inhibition of PKCα-Dependent NADPH Oxidase/ROS and NF-κB"

Article Title: HO-1 Induction by CO-RM2 Attenuates TNF-α-Induced Cytosolic Phospholipase A2 Expression via Inhibition of PKCα-Dependent NADPH Oxidase/ROS and NF-κB

Journal: Mediators of Inflammation

doi: 10.1155/2014/279171

Effect of CORM-2 on TNF- α -induced NF- κ B activation and cPLA 2 expression. (a) Cells were pretreated with CORM-2 or iCORM-2 for 16 h and then incubated with TNF- α for 2 h. Chromatin immunoprecipitation (ChIP) assays were performed using an anti-p65 antibody. (b) Cells were transfected with NF- κ B-luc reporter gene, pretreated with or without CORM-2 for 16 h, and then incubated with TNF- α for 4 h. NF- κ B promoter activity was determined. ((c), (d), and (f)) Cells were pretreated with or without CORM-2 or iCORM-2 for 16 h and then treated with TNF- α for the indicated time intervals. The levels of phospho-p65, phospho-IKK α / β , phospho-JNK1/2, phospho-p38 MAPK, and HO-1 were determined by Western blotting. (e) Cells were pretreated with or without CORM-2 for 16 h and then incubated with TNF- α for 90 min. The cells were added with (5 μ M) DHE and images acquired to determine the ROS generation using a fluorescence microscopy. All analyses were performed on samples from 4 RA patients. (f) Immunohistochemical staining for HO-1, cPLA 2 , or vimentin and hematoxylin and eosin (H E) staining of serial sections of ankle joints from mice injected with phosphate-buffered saline (sham) ((A)–(E)), mice injected with TNF- α ((F)–(J)), and mice injected with CORM-2 for 16 h followed by TNF- α for 24 h ((K)–(O)) were performed. Arrowheads indicate positive staining. Results are representative of 3 mice per experimental group. Results are representative of 3 independent experiments. In (a), (b), (c), and (g), values are the mean ± SEM. * P
Figure Legend Snippet: Effect of CORM-2 on TNF- α -induced NF- κ B activation and cPLA 2 expression. (a) Cells were pretreated with CORM-2 or iCORM-2 for 16 h and then incubated with TNF- α for 2 h. Chromatin immunoprecipitation (ChIP) assays were performed using an anti-p65 antibody. (b) Cells were transfected with NF- κ B-luc reporter gene, pretreated with or without CORM-2 for 16 h, and then incubated with TNF- α for 4 h. NF- κ B promoter activity was determined. ((c), (d), and (f)) Cells were pretreated with or without CORM-2 or iCORM-2 for 16 h and then treated with TNF- α for the indicated time intervals. The levels of phospho-p65, phospho-IKK α / β , phospho-JNK1/2, phospho-p38 MAPK, and HO-1 were determined by Western blotting. (e) Cells were pretreated with or without CORM-2 for 16 h and then incubated with TNF- α for 90 min. The cells were added with (5 μ M) DHE and images acquired to determine the ROS generation using a fluorescence microscopy. All analyses were performed on samples from 4 RA patients. (f) Immunohistochemical staining for HO-1, cPLA 2 , or vimentin and hematoxylin and eosin (H E) staining of serial sections of ankle joints from mice injected with phosphate-buffered saline (sham) ((A)–(E)), mice injected with TNF- α ((F)–(J)), and mice injected with CORM-2 for 16 h followed by TNF- α for 24 h ((K)–(O)) were performed. Arrowheads indicate positive staining. Results are representative of 3 mice per experimental group. Results are representative of 3 independent experiments. In (a), (b), (c), and (g), values are the mean ± SEM. * P

Techniques Used: Activation Assay, Expressing, Incubation, Chromatin Immunoprecipitation, Transfection, Activity Assay, Western Blot, Fluorescence, Microscopy, Immunohistochemistry, Staining, Mouse Assay, Injection

CORM-2 enhances HO-1 expression and modulates TNF- α -induced cPLA 2 expression in RASFs. RASFs were treated with (a) different concentrations or (b) 25 μ M of CORM-2 for the indicated time intervals. (c) RASFs were incubated with various concentrations of TNF- α for the indicated time intervals. (d) Cells were incubated with TNF- α (30 ng/mL) for the indicated time intervals. The mRNA levels of cPLA 2 and cPLA 2 -Luc promoter were measured. (e) Cells were pretreated with different concentrations of CORM-2 for 16 h and then incubated with TNF- α for 16 h. Cells were pretreated with (f) CORM-2 or iCORM-2 or (h) were transfected with scrambled (scrb) or HO-1 siRNA and then incubated without or with TNF- α for 16 h. ((a), (c), (e), (f), and (h)) The cell lysates were analyzed by Western blotting using an anti-cPLA 2 , anti-HO-1, or anti-GAPDH (control). ((g), open bars) RASFs were pretreated without or with CORM-2, or iCORM-2 for 16 h, and incubated with TNF- α (30 ng/mL) for 6 h. cPLA 2 mRNA was analyzed by quantitative real-time PCR. ((g), shaded bars) RASFs were transfected with a cPLA 2 -Luc reporter gene, pretreated without or with CORM-2 or iCORM-2 for 16 h, and incubated with TNF- α (30 ng/mL) for 6 h. Promoter luciferase activity was analyzed. All analyses were performed on samples from 4 RA patients. Results are representative of 3 independent experiments. Values are the mean ± SEM. ((b)–(d)) * P
Figure Legend Snippet: CORM-2 enhances HO-1 expression and modulates TNF- α -induced cPLA 2 expression in RASFs. RASFs were treated with (a) different concentrations or (b) 25 μ M of CORM-2 for the indicated time intervals. (c) RASFs were incubated with various concentrations of TNF- α for the indicated time intervals. (d) Cells were incubated with TNF- α (30 ng/mL) for the indicated time intervals. The mRNA levels of cPLA 2 and cPLA 2 -Luc promoter were measured. (e) Cells were pretreated with different concentrations of CORM-2 for 16 h and then incubated with TNF- α for 16 h. Cells were pretreated with (f) CORM-2 or iCORM-2 or (h) were transfected with scrambled (scrb) or HO-1 siRNA and then incubated without or with TNF- α for 16 h. ((a), (c), (e), (f), and (h)) The cell lysates were analyzed by Western blotting using an anti-cPLA 2 , anti-HO-1, or anti-GAPDH (control). ((g), open bars) RASFs were pretreated without or with CORM-2, or iCORM-2 for 16 h, and incubated with TNF- α (30 ng/mL) for 6 h. cPLA 2 mRNA was analyzed by quantitative real-time PCR. ((g), shaded bars) RASFs were transfected with a cPLA 2 -Luc reporter gene, pretreated without or with CORM-2 or iCORM-2 for 16 h, and incubated with TNF- α (30 ng/mL) for 6 h. Promoter luciferase activity was analyzed. All analyses were performed on samples from 4 RA patients. Results are representative of 3 independent experiments. Values are the mean ± SEM. ((b)–(d)) * P

Techniques Used: Expressing, Incubation, Transfection, Western Blot, Real-time Polymerase Chain Reaction, Luciferase, Activity Assay

18) Product Images from "Antioxidant and Anti-Inflammatory Effects of Herbal Formula SC-E3 in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages"

Article Title: Antioxidant and Anti-Inflammatory Effects of Herbal Formula SC-E3 in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2017/1725246

Effects of SC-E3 on the Nrf2/HO-1 signaling pathway in RAW 264.7 macrophages. (a) Induction of HO-1 by SC-E3. Cells were treated with different concentrations of SC-E3 (50, 100, 300, or 500 μ g/mL) for 18 h. (b) Cells were treated with 300 μ g/mL SC-E3 for the indicated times. (c) Nuclear accumulation of Nrf2 by SC-E3. Nrf2 was immunoblotted in the nuclear fractions of cells treated with 300 μ g/mL of SC-E3 for the indicated times. (d) Immunofluorescence images of the nuclear translocation of Nrf2 induced by SC-E3. RAW 264.7 cells were treated with 300 μ g/mL of SC-E3 for 3 h. (e) Blocking of the inhibitory effect of SC-E3 on LPS-induced NO production by SnPP (an HO-1 inhibitor). RAW 264.7 cells were pretreated with SC-E3 (300 μ g/mL) for 1 h in the presence or absence of SnPP (50 nM, 30 min) and then stimulated with LPS (1 μ g/mL) for 18 h. ∗∗∗ p
Figure Legend Snippet: Effects of SC-E3 on the Nrf2/HO-1 signaling pathway in RAW 264.7 macrophages. (a) Induction of HO-1 by SC-E3. Cells were treated with different concentrations of SC-E3 (50, 100, 300, or 500 μ g/mL) for 18 h. (b) Cells were treated with 300 μ g/mL SC-E3 for the indicated times. (c) Nuclear accumulation of Nrf2 by SC-E3. Nrf2 was immunoblotted in the nuclear fractions of cells treated with 300 μ g/mL of SC-E3 for the indicated times. (d) Immunofluorescence images of the nuclear translocation of Nrf2 induced by SC-E3. RAW 264.7 cells were treated with 300 μ g/mL of SC-E3 for 3 h. (e) Blocking of the inhibitory effect of SC-E3 on LPS-induced NO production by SnPP (an HO-1 inhibitor). RAW 264.7 cells were pretreated with SC-E3 (300 μ g/mL) for 1 h in the presence or absence of SnPP (50 nM, 30 min) and then stimulated with LPS (1 μ g/mL) for 18 h. ∗∗∗ p

Techniques Used: Immunofluorescence, Translocation Assay, Blocking Assay

19) Product Images from "The Food Contaminants Nivalenol and Deoxynivalenol Induce Inflammation in Intestinal Epithelial Cells by Regulating Reactive Oxygen Species Release"

Article Title: The Food Contaminants Nivalenol and Deoxynivalenol Induce Inflammation in Intestinal Epithelial Cells by Regulating Reactive Oxygen Species Release

Journal: Nutrients

doi: 10.3390/nu9121343

( A ) Effect of NIV, DON, and their combination (NIV + DON; 5 μM), on Nrf2 nuclear translocation, evaluated using immunofluorescence assay confocal microscopy. Scale bar: 10 μm. Blue and green fluorescences indicate localization of nucleus (DAPI) and Nrf2, respectively; ( B ) Effect of NIV; ( C ) DON; and ( D ) their combination (NIV + DON) on HO-1 expression in the IEC-6 cells, evaluated by cytofluorimetric technique. Values, mean ± s.e.m., are expressed as mean fluorescence intensity. ° Denotes p
Figure Legend Snippet: ( A ) Effect of NIV, DON, and their combination (NIV + DON; 5 μM), on Nrf2 nuclear translocation, evaluated using immunofluorescence assay confocal microscopy. Scale bar: 10 μm. Blue and green fluorescences indicate localization of nucleus (DAPI) and Nrf2, respectively; ( B ) Effect of NIV; ( C ) DON; and ( D ) their combination (NIV + DON) on HO-1 expression in the IEC-6 cells, evaluated by cytofluorimetric technique. Values, mean ± s.e.m., are expressed as mean fluorescence intensity. ° Denotes p

Techniques Used: Translocation Assay, Immunofluorescence, Confocal Microscopy, Expressing, Fluorescence

20) Product Images from "The Neuroprotective Effect of Dimethyl Fumarate in an MPTP-Mouse Model of Parkinson's Disease: Involvement of Reactive Oxygen Species/Nuclear Factor-κB/Nuclear Transcription Factor Related to NF-E2"

Article Title: The Neuroprotective Effect of Dimethyl Fumarate in an MPTP-Mouse Model of Parkinson's Disease: Involvement of Reactive Oxygen Species/Nuclear Factor-κB/Nuclear Transcription Factor Related to NF-E2

Journal: Antioxidants & Redox Signaling

doi: 10.1089/ars.2016.6800

Effects of DMF treatment on antioxidant response activation. The expression of Nrf-2 levels showed a tendency to decrease after MPTP administration as compared with control mice (A ) . DMF treatment caused a dose-dependent upregulation of Nrf-2 levels (A ) . In terms of antioxidant enzyme expression in MPTP-injured mice, there was a greater decrease in HO-1 level as compared with Mn-SOD (C , B ) . Treatment with 30 mg/kg DMF increased Mn-SOD expression (B ) ; DMF, at 10 mg/kg, was ineffective (B ) . In contrast, the lower DMF dose (10 mg/kg) increased HO-1 expression (C ) but not significantly, compared with DMF treatment at 30 mg/kg that returned HO-1 expression to control values (C ) . MPTP-injected mice had an increased nNOS expression (D ) , and DMF treatment (especially at 30 mg/kg) reduced nNOS levels as compared with sham (D ) . Furthermore, immunohistochemical analysis revealed positive NT immunostaining in MPTP-injected mice after 8 days (F, F1, I) compared with control mice (E, E1, I) , and DMF limited this effect in a dose-dependent manner (G, G1 , H, H1 , respectively, and I) . In addition, the decline in GSH/GSSG ratio after MPTP intoxication was almost completely prevented by 30 mg/kg DMF (J) . Data are means ± SD of 10 mice for each group. *** p
Figure Legend Snippet: Effects of DMF treatment on antioxidant response activation. The expression of Nrf-2 levels showed a tendency to decrease after MPTP administration as compared with control mice (A ) . DMF treatment caused a dose-dependent upregulation of Nrf-2 levels (A ) . In terms of antioxidant enzyme expression in MPTP-injured mice, there was a greater decrease in HO-1 level as compared with Mn-SOD (C , B ) . Treatment with 30 mg/kg DMF increased Mn-SOD expression (B ) ; DMF, at 10 mg/kg, was ineffective (B ) . In contrast, the lower DMF dose (10 mg/kg) increased HO-1 expression (C ) but not significantly, compared with DMF treatment at 30 mg/kg that returned HO-1 expression to control values (C ) . MPTP-injected mice had an increased nNOS expression (D ) , and DMF treatment (especially at 30 mg/kg) reduced nNOS levels as compared with sham (D ) . Furthermore, immunohistochemical analysis revealed positive NT immunostaining in MPTP-injected mice after 8 days (F, F1, I) compared with control mice (E, E1, I) , and DMF limited this effect in a dose-dependent manner (G, G1 , H, H1 , respectively, and I) . In addition, the decline in GSH/GSSG ratio after MPTP intoxication was almost completely prevented by 30 mg/kg DMF (J) . Data are means ± SD of 10 mice for each group. *** p

Techniques Used: Activation Assay, Expressing, Mouse Assay, Injection, Immunohistochemistry, Immunostaining

21) Product Images from "Nimbolide protects against endotoxin-induced acute respiratory distress syndrome by inhibiting TNF-α mediated NF-κB and HDAC-3 nuclear translocation"

Article Title: Nimbolide protects against endotoxin-induced acute respiratory distress syndrome by inhibiting TNF-α mediated NF-κB and HDAC-3 nuclear translocation

Journal: Cell Death & Disease

doi: 10.1038/s41419-018-1247-9

Nimbolide protects the lungs from nitrosative and oxidative stress. Cells were pre-treated with nimbolide for 24 h and oxidative stress was induced with LPS (1 µg/ml). After 24 h of incubation, the nitrite levels were measured in cell culture supernatants of ( a ) RAW 264.7 and ( b ) differentiated THP-1 cells by Griess assay. Whole cell protein was extracted and subjected to western blot and measured the expression of nitrotyrosine and iNOS in both ( c ) RAW 264.7 and ( d ) differentiated THP-1 cells. After 30 min of LPS stimulation, intracellular and mitochondrial ROS (mROS) levels were measured by ( e , g ) DCFDA and ( f , h ) MitoSOX Red staining in both RAW 264.7 and differentiated THP-1 cells, respectively. The fluorescent images were captured at ×200 magnification from different groups. ( i ) RAW264.7 cells were pre-treated with nimbolide (1 µM) and oxidative stress was induced by LPS (1 µg/ml). After 12 h of LPS exposure, cells were incubated with Pa and macrophage bactericidal activity was calculated by counting Pa colonies on nutrient agar plates. Western blot was performed to determine the Nrf-2, HO-1, and SOD1 expression in both ( j ) RAW 264.7 and ( k ) differentiated THP-1 cells from whole cell lysate. Data represented as mean ± SEM ( n = 3 independent experiments). * P
Figure Legend Snippet: Nimbolide protects the lungs from nitrosative and oxidative stress. Cells were pre-treated with nimbolide for 24 h and oxidative stress was induced with LPS (1 µg/ml). After 24 h of incubation, the nitrite levels were measured in cell culture supernatants of ( a ) RAW 264.7 and ( b ) differentiated THP-1 cells by Griess assay. Whole cell protein was extracted and subjected to western blot and measured the expression of nitrotyrosine and iNOS in both ( c ) RAW 264.7 and ( d ) differentiated THP-1 cells. After 30 min of LPS stimulation, intracellular and mitochondrial ROS (mROS) levels were measured by ( e , g ) DCFDA and ( f , h ) MitoSOX Red staining in both RAW 264.7 and differentiated THP-1 cells, respectively. The fluorescent images were captured at ×200 magnification from different groups. ( i ) RAW264.7 cells were pre-treated with nimbolide (1 µM) and oxidative stress was induced by LPS (1 µg/ml). After 12 h of LPS exposure, cells were incubated with Pa and macrophage bactericidal activity was calculated by counting Pa colonies on nutrient agar plates. Western blot was performed to determine the Nrf-2, HO-1, and SOD1 expression in both ( j ) RAW 264.7 and ( k ) differentiated THP-1 cells from whole cell lysate. Data represented as mean ± SEM ( n = 3 independent experiments). * P

Techniques Used: Incubation, Cell Culture, Griess Assay, Western Blot, Expressing, Staining, Activity Assay

Nimbolide modulates the pro- and anti-inflammatory cytokines and chemokines. The proinflammatory cytokines and chemokines (IL-6, IL-12 (p40), MIP-1α, MIP-1β, and TNF-α) and anti-inflammatory cytokines (IL-4, IL-10, and IL-13) expression were determined by multiplex and ELISA. a – h Animal lung tissue lysate was prepared and evaluated for the levels of aforementioned inflammatory cytokines and chemokines by multiplex. i , j lung tissue supernatants were subjected to ELISA to determine IL-1β and TGF-β cytokines expression. k Griess assay was performed to determine the nitrite levels in cell lysate of lung tissues. l The antioxidant GSH levels were measured in lung tissue. m The MPO as well as ( n ) iNOS, nitrotyrosine, Nrf-2, HO-1, and SOD-1 protein expressions were determined by immunoblot analysis. Data presented as mean ± SEM (n=8 mice per group). * P
Figure Legend Snippet: Nimbolide modulates the pro- and anti-inflammatory cytokines and chemokines. The proinflammatory cytokines and chemokines (IL-6, IL-12 (p40), MIP-1α, MIP-1β, and TNF-α) and anti-inflammatory cytokines (IL-4, IL-10, and IL-13) expression were determined by multiplex and ELISA. a – h Animal lung tissue lysate was prepared and evaluated for the levels of aforementioned inflammatory cytokines and chemokines by multiplex. i , j lung tissue supernatants were subjected to ELISA to determine IL-1β and TGF-β cytokines expression. k Griess assay was performed to determine the nitrite levels in cell lysate of lung tissues. l The antioxidant GSH levels were measured in lung tissue. m The MPO as well as ( n ) iNOS, nitrotyrosine, Nrf-2, HO-1, and SOD-1 protein expressions were determined by immunoblot analysis. Data presented as mean ± SEM (n=8 mice per group). * P

Techniques Used: Expressing, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Griess Assay, Mouse Assay

22) Product Images from "HO-1 Induction by Selaginella tamariscina Extract Inhibits Inflammatory Response in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages"

Article Title: HO-1 Induction by Selaginella tamariscina Extract Inhibits Inflammatory Response in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2018/7816923

Induction of Nrf2/HO-1 signaling pathway by STE. (a) Immunofluorescence images of the nuclear translocation of Nrf2 induced by STE. RAW 264.7 cells were treated with STE (300 μ g/mL) for 3 h. (b) Induction of HO-1 by STE. RAW 264.7 cells were treated with various concentrations of STE (10, 100, or 300 μ g/mL) for 18 h, lysed, and western blotted. (Significant vs. vehicle-treated control, ∗ p
Figure Legend Snippet: Induction of Nrf2/HO-1 signaling pathway by STE. (a) Immunofluorescence images of the nuclear translocation of Nrf2 induced by STE. RAW 264.7 cells were treated with STE (300 μ g/mL) for 3 h. (b) Induction of HO-1 by STE. RAW 264.7 cells were treated with various concentrations of STE (10, 100, or 300 μ g/mL) for 18 h, lysed, and western blotted. (Significant vs. vehicle-treated control, ∗ p

Techniques Used: Immunofluorescence, Translocation Assay, Western Blot

23) Product Images from "Cafestol Inhibits Cyclic-Strain-Induced Interleukin-8, Intercellular Adhesion Molecule-1, and Monocyte Chemoattractant Protein-1 Production in Vascular Endothelial Cells"

Article Title: Cafestol Inhibits Cyclic-Strain-Induced Interleukin-8, Intercellular Adhesion Molecule-1, and Monocyte Chemoattractant Protein-1 Production in Vascular Endothelial Cells

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2018/7861518

Effects of cafestol on HO-1 expression in the presence of cyclic strain treatment. HUVECs were treated with cafestol 12 h prior to cyclic strain treatment for 12 h. (a) The mRNA level of HO-1 was analyzed through qPCR and normalized to GAPDH. (b) Immunoblotting of HO-1 and GAPDH was performed, and the bands were quantitated using ImageJ. The data represent the mean ± SEM of three independent experiments. ∗ P
Figure Legend Snippet: Effects of cafestol on HO-1 expression in the presence of cyclic strain treatment. HUVECs were treated with cafestol 12 h prior to cyclic strain treatment for 12 h. (a) The mRNA level of HO-1 was analyzed through qPCR and normalized to GAPDH. (b) Immunoblotting of HO-1 and GAPDH was performed, and the bands were quantitated using ImageJ. The data represent the mean ± SEM of three independent experiments. ∗ P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

24) Product Images from "Inhibitory Effects of Momordicine I on High-Glucose-Induced Cell Proliferation and Collagen Synthesis in Rat Cardiac Fibroblasts"

Article Title: Inhibitory Effects of Momordicine I on High-Glucose-Induced Cell Proliferation and Collagen Synthesis in Rat Cardiac Fibroblasts

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2018/3939714

Momordicine I increases Nrf2 translocation and HO-1 protein expression in rat cardiac fibroblasts. (a) Effect of momordicine I on Nrf2 translocation. Cells were treated with or without momordicine I (1 μ M) for 12 h, followed by a normal-glucose medium or high-glucose medium for 12 h. N: nuclear extract. Results are presented as the mean ± SEM ( n = 4). (b) HO-1 expression was determined through Western blotting. Cells were pretreated with momordicine I (1 μ M) or not for 12 h, followed by the control medium or high-glucose medium for 24 h. Results are presented as the mean ± SEM ( n = 3). The relative protein expression of nuclear Nrf2 to PARP and HO-1 to GAPDH are presented in the bar graphs. ∗ P
Figure Legend Snippet: Momordicine I increases Nrf2 translocation and HO-1 protein expression in rat cardiac fibroblasts. (a) Effect of momordicine I on Nrf2 translocation. Cells were treated with or without momordicine I (1 μ M) for 12 h, followed by a normal-glucose medium or high-glucose medium for 12 h. N: nuclear extract. Results are presented as the mean ± SEM ( n = 4). (b) HO-1 expression was determined through Western blotting. Cells were pretreated with momordicine I (1 μ M) or not for 12 h, followed by the control medium or high-glucose medium for 24 h. Results are presented as the mean ± SEM ( n = 3). The relative protein expression of nuclear Nrf2 to PARP and HO-1 to GAPDH are presented in the bar graphs. ∗ P

Techniques Used: Translocation Assay, Expressing, Western Blot

25) Product Images from "Cafestol Inhibits Cyclic-Strain-Induced Interleukin-8, Intercellular Adhesion Molecule-1, and Monocyte Chemoattractant Protein-1 Production in Vascular Endothelial Cells"

Article Title: Cafestol Inhibits Cyclic-Strain-Induced Interleukin-8, Intercellular Adhesion Molecule-1, and Monocyte Chemoattractant Protein-1 Production in Vascular Endothelial Cells

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2018/7861518

Effects of cafestol on HO-1 expression in the presence of cyclic strain treatment. HUVECs were treated with cafestol 12 h prior to cyclic strain treatment for 12 h. (a) The mRNA level of HO-1 was analyzed through qPCR and normalized to GAPDH. (b) Immunoblotting of HO-1 and GAPDH was performed, and the bands were quantitated using ImageJ. The data represent the mean ± SEM of three independent experiments. ∗ P
Figure Legend Snippet: Effects of cafestol on HO-1 expression in the presence of cyclic strain treatment. HUVECs were treated with cafestol 12 h prior to cyclic strain treatment for 12 h. (a) The mRNA level of HO-1 was analyzed through qPCR and normalized to GAPDH. (b) Immunoblotting of HO-1 and GAPDH was performed, and the bands were quantitated using ImageJ. The data represent the mean ± SEM of three independent experiments. ∗ P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

26) Product Images from "Nutritional Mushroom Treatment in Meniere’s Disease with Coriolus versicolor: A Rationale for Therapeutic Intervention in Neuroinflammation and Antineurodegeneration"

Article Title: Nutritional Mushroom Treatment in Meniere’s Disease with Coriolus versicolor: A Rationale for Therapeutic Intervention in Neuroinflammation and Antineurodegeneration

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms21010284

Heme oxygenase-1 levels in lymphocytes and in plasma from MD patients. Samples from MD patients were assayed for heme oxygenase-1 (HO-1) by western blot as described in Materials and Methods. A representative immunoblot is shown. β-actin has been used as loading control ( c , d ). The bar graph shows the densitometric evaluation ( a , b ) and values are expressed as mean ± SEM of independent analyses on 22 patients (MD plus Coriolus biomass) and, respectively, on 18 patients (MD alone), per group. * p
Figure Legend Snippet: Heme oxygenase-1 levels in lymphocytes and in plasma from MD patients. Samples from MD patients were assayed for heme oxygenase-1 (HO-1) by western blot as described in Materials and Methods. A representative immunoblot is shown. β-actin has been used as loading control ( c , d ). The bar graph shows the densitometric evaluation ( a , b ) and values are expressed as mean ± SEM of independent analyses on 22 patients (MD plus Coriolus biomass) and, respectively, on 18 patients (MD alone), per group. * p

Techniques Used: Western Blot

27) Product Images from "Acerogenin A from Acer nikoense Maxim Prevents Oxidative Stress-Induced Neuronal Cell Death through Nrf2-Mediated Heme Oxygenase-1 Expression in Mouse Hippocampal HT22 Cell Line"

Article Title: Acerogenin A from Acer nikoense Maxim Prevents Oxidative Stress-Induced Neuronal Cell Death through Nrf2-Mediated Heme Oxygenase-1 Expression in Mouse Hippocampal HT22 Cell Line

Journal: Molecules

doi: 10.3390/molecules200712545

Effects of acerogenin A-induced HO-1 expression through the phosphatidylinositol 3-kinase (PI3K)/AKT cascade. HT22 cells were treated with acerogenin A (30 μM) for the indicated times ( A ). HT22 cells were pre-incubated with or without 10 μM LY294002 for 1 h and then incubated in the absence or presence of 20 μM of acerogenin A for 60 min (p-AKT) or 12 h (HO-1) ( B ). HT22 cells untreated or treated with acerogenin A (30 μM) in the presence or absence of LY294002 (10 μM) for 12 h were exposed to 5 mM glutamate for 12 h ( C ). Western blot analysis was performed, and representative blots of three independent experiments are shown. Data are presented as the mean ± SD values of three independent experiments. * p
Figure Legend Snippet: Effects of acerogenin A-induced HO-1 expression through the phosphatidylinositol 3-kinase (PI3K)/AKT cascade. HT22 cells were treated with acerogenin A (30 μM) for the indicated times ( A ). HT22 cells were pre-incubated with or without 10 μM LY294002 for 1 h and then incubated in the absence or presence of 20 μM of acerogenin A for 60 min (p-AKT) or 12 h (HO-1) ( B ). HT22 cells untreated or treated with acerogenin A (30 μM) in the presence or absence of LY294002 (10 μM) for 12 h were exposed to 5 mM glutamate for 12 h ( C ). Western blot analysis was performed, and representative blots of three independent experiments are shown. Data are presented as the mean ± SD values of three independent experiments. * p

Techniques Used: Expressing, Incubation, Western Blot

The effects of acerogenin A on the nuclear translocation of Nrf2 ( A ) and Nrf2-mediated HO-1 ( B ) in HT22 cells. HT22 cells were treated with 30 μM of acerogenin A for 0, 30, 60, and 90 min ( A ). The nuclei were fractionated from the cytosol by using PER-Mammalian Protein Extraction Buffer, as described in the Experimental Section ( A ). HT22 cells were transiently transfected with Nrf2 siRNA and then treated with 30 μM acerogenin A for 12 h ( B ). Western blot analysis was performed, and representative blots of three independent experiments are shown. Data are presented as the mean value ± SD values of three independent experiments. * p
Figure Legend Snippet: The effects of acerogenin A on the nuclear translocation of Nrf2 ( A ) and Nrf2-mediated HO-1 ( B ) in HT22 cells. HT22 cells were treated with 30 μM of acerogenin A for 0, 30, 60, and 90 min ( A ). The nuclei were fractionated from the cytosol by using PER-Mammalian Protein Extraction Buffer, as described in the Experimental Section ( A ). HT22 cells were transiently transfected with Nrf2 siRNA and then treated with 30 μM acerogenin A for 12 h ( B ). Western blot analysis was performed, and representative blots of three independent experiments are shown. Data are presented as the mean value ± SD values of three independent experiments. * p

Techniques Used: Translocation Assay, Protein Extraction, Transfection, Western Blot

The effects of acerogenin A on heme oxygenase (HO)-1 expression ( A , B ) and acerogenin A-induced HO-1 on glutamate-induced oxidative neurotoxicity ( C ) and ROS generation ( D ). HT22 cells were incubated with indicated concentrations of acerogenin A for 12 h ( A , B ). HT22 cells were treated with 30 μM of acerogenin A in the presence or absence of 50 μM SnPP IX and HO-1 siRNA, and then exposed to glutamate (5 mM) for 12 h ( C , D ). Exposure of HT22 cells to 5 mM glutamate for 12 h to increase ROS production, followed by incubation with 10 μM of the ROS-sensitive fluorophore dichlorofluorescein (DCF) ( D ). Western blot analysis was performed, and representative blots of three independent experiments are shown. Data are presented as the mean ± SD values of three independent experiments. * p
Figure Legend Snippet: The effects of acerogenin A on heme oxygenase (HO)-1 expression ( A , B ) and acerogenin A-induced HO-1 on glutamate-induced oxidative neurotoxicity ( C ) and ROS generation ( D ). HT22 cells were incubated with indicated concentrations of acerogenin A for 12 h ( A , B ). HT22 cells were treated with 30 μM of acerogenin A in the presence or absence of 50 μM SnPP IX and HO-1 siRNA, and then exposed to glutamate (5 mM) for 12 h ( C , D ). Exposure of HT22 cells to 5 mM glutamate for 12 h to increase ROS production, followed by incubation with 10 μM of the ROS-sensitive fluorophore dichlorofluorescein (DCF) ( D ). Western blot analysis was performed, and representative blots of three independent experiments are shown. Data are presented as the mean ± SD values of three independent experiments. * p

Techniques Used: Expressing, Incubation, Western Blot

28) Product Images from "Fenofibrate exerts protective effects against gentamicin-induced toxicity in cochlear hair cells by activating antioxidant enzymes"

Article Title: Fenofibrate exerts protective effects against gentamicin-induced toxicity in cochlear hair cells by activating antioxidant enzymes

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2017.2916

Effect of fenofibrate on the expression of antioxidant enzymes in rat cochlea. (A) Cochlear explants were treated with medium alone, GM (300 µ M) for 18 h, fenofibrate (100 µ M) pre-treated for 4 h and then co-treated with GM (300 µ M) for 18 h, and fenofibrate (100 µ M) only for 22 h. The organ of Corti was collected and proteins were extracted. Total cell lysates were separated by 10% SDS-PAGE to detect PPAR-α, catalase, SOD-1 and HO-1 proteins. The protein and mRNA levels of β-actin were determined as controls. (B) The inner ears from SD rat [saline group (control), intraperitoneally injected with 200 mg/kg GM for 4 days (GM), 100 mg/kg fenofibrate for 10 days followed by GM (FF + GM) and fenofibrate alone (FF)] were removed and embedded in paraffin. Next, 4- µ m-thick sections were prepared. For immunohistochemistry studies, a commercial kit (LSAB Universal K680) was used to detect the expression levels of catalase, SOD-1 and HO-1 in the cochlear duct regions. GM, gentamicin; FF, fenofibrate; PPAR, peroxisome proliferator-activated receptor; SOD-1, superoxide dismutase-1; HO-1, heme oxygenase-1; SV, stria vascularis; SLig, spiral limbus; SLim, spiral ligament.
Figure Legend Snippet: Effect of fenofibrate on the expression of antioxidant enzymes in rat cochlea. (A) Cochlear explants were treated with medium alone, GM (300 µ M) for 18 h, fenofibrate (100 µ M) pre-treated for 4 h and then co-treated with GM (300 µ M) for 18 h, and fenofibrate (100 µ M) only for 22 h. The organ of Corti was collected and proteins were extracted. Total cell lysates were separated by 10% SDS-PAGE to detect PPAR-α, catalase, SOD-1 and HO-1 proteins. The protein and mRNA levels of β-actin were determined as controls. (B) The inner ears from SD rat [saline group (control), intraperitoneally injected with 200 mg/kg GM for 4 days (GM), 100 mg/kg fenofibrate for 10 days followed by GM (FF + GM) and fenofibrate alone (FF)] were removed and embedded in paraffin. Next, 4- µ m-thick sections were prepared. For immunohistochemistry studies, a commercial kit (LSAB Universal K680) was used to detect the expression levels of catalase, SOD-1 and HO-1 in the cochlear duct regions. GM, gentamicin; FF, fenofibrate; PPAR, peroxisome proliferator-activated receptor; SOD-1, superoxide dismutase-1; HO-1, heme oxygenase-1; SV, stria vascularis; SLig, spiral limbus; SLim, spiral ligament.

Techniques Used: Expressing, SDS Page, Injection, Immunohistochemistry

Effect of SnPPIX, a HO-1 inhibitor, on fenofibrate-mediated protection of zebrafish neuromasts. (A) The neuromasts of zebrafish were (50 µ M) for 1 h (GM, panels b and g), pre-treated with fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (500 µ M) for 1 h (FF + GM, panels c and h), pre-treated with SnPPIX (10 µ M) and fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (50 µ M) for 1 h (SnPP + FF + GM, panels d and i), and fenofibrate alone (FF, panels e and j). One of the occipital neuromasts (OC1) is shown in panels a-e, and a posterior neuromast is (P1) shown in panels f-j. (B) Quantitative analysis of neuromast survival. Histogram represents the mean viability of neuromasts. Zebrafish were treated with medium alone (cont), GM (50 µ M) for 1 h (OC and P1: 30.7±14.17, p≤ 0.001 and 24.2±9.29, p≤0.0002, respectively), pre-treated with fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (500 µ M) for 1 h (OC and P1: 69.3±19.2, p≤0.003 and 72.5±17.86, p≤0.0006, respectively), pre-treated with SnPPIX (10 µ M) and fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (50 µ M) for 1 h (OC and P1: 38.6±10.00, p≤0.001 and 40.0±10.00, p≤0.002, respectively), and fenofibrate alone (FF). # p
Figure Legend Snippet: Effect of SnPPIX, a HO-1 inhibitor, on fenofibrate-mediated protection of zebrafish neuromasts. (A) The neuromasts of zebrafish were (50 µ M) for 1 h (GM, panels b and g), pre-treated with fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (500 µ M) for 1 h (FF + GM, panels c and h), pre-treated with SnPPIX (10 µ M) and fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (50 µ M) for 1 h (SnPP + FF + GM, panels d and i), and fenofibrate alone (FF, panels e and j). One of the occipital neuromasts (OC1) is shown in panels a-e, and a posterior neuromast is (P1) shown in panels f-j. (B) Quantitative analysis of neuromast survival. Histogram represents the mean viability of neuromasts. Zebrafish were treated with medium alone (cont), GM (50 µ M) for 1 h (OC and P1: 30.7±14.17, p≤ 0.001 and 24.2±9.29, p≤0.0002, respectively), pre-treated with fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (500 µ M) for 1 h (OC and P1: 69.3±19.2, p≤0.003 and 72.5±17.86, p≤0.0006, respectively), pre-treated with SnPPIX (10 µ M) and fenofibrate (10 µ M) for 0.5 h and then co-treated with GM (50 µ M) for 1 h (OC and P1: 38.6±10.00, p≤0.001 and 40.0±10.00, p≤0.002, respectively), and fenofibrate alone (FF). # p

Techniques Used:

Effect of SnPPIX, an HO-1 inhibitor, on fenofibrate-mediated protection of the sensory hair cells. (A) Sensory hair cells were stained with phalloidin-TRITC and observed under a fluorescence microscope. Cochlear explants were treated with medium alone, GM (300 µ M) for 24 h (32.7±3.78, p≤0.00001), pre-treated with fenofibrate (100 µ M) for 4 h and then co-treated with GM (300 µ M) for 24 h (83.8±3.78, p≤0.00006), and pre-treated with SnPPIX (10 µ M) and fenofibrate (100 µ M) for 4 h and then further incubated with GM (300 µ M) for 24 h (59.3±3.78, p≤0.001). (B) Quantitative analysis of the survival of sensory hair cells. Histogram represents mean hair cell viability. * p
Figure Legend Snippet: Effect of SnPPIX, an HO-1 inhibitor, on fenofibrate-mediated protection of the sensory hair cells. (A) Sensory hair cells were stained with phalloidin-TRITC and observed under a fluorescence microscope. Cochlear explants were treated with medium alone, GM (300 µ M) for 24 h (32.7±3.78, p≤0.00001), pre-treated with fenofibrate (100 µ M) for 4 h and then co-treated with GM (300 µ M) for 24 h (83.8±3.78, p≤0.00006), and pre-treated with SnPPIX (10 µ M) and fenofibrate (100 µ M) for 4 h and then further incubated with GM (300 µ M) for 24 h (59.3±3.78, p≤0.001). (B) Quantitative analysis of the survival of sensory hair cells. Histogram represents mean hair cell viability. * p

Techniques Used: Staining, Fluorescence, Microscopy, Incubation

29) Product Images from "Neuroprotective Effect of β-Lapachone in MPTP-Induced Parkinson’s Disease Mouse Model: Involvement of Astroglial p-AMPK/Nrf2/HO-1 Signaling Pathways"

Article Title: Neuroprotective Effect of β-Lapachone in MPTP-Induced Parkinson’s Disease Mouse Model: Involvement of Astroglial p-AMPK/Nrf2/HO-1 Signaling Pathways

Journal: Biomolecules & Therapeutics

doi: 10.4062/biomolther.2018.234

β-LAP upregulated the expression of HO-1 in striatal astrocytes. (A) Quantitative real-time PCR data showing the mRNA expression of HO-1 in striatum of MPTP-injected mice (n=5 per group). (B) Western blot analysis showing the expression of HO-1 protein in the striatum of MPTP-injected mice (n=5 per group). The quantification data are provided in the right panel. (C) The double IF staining results indicate that the HO-1 protein (green) was upregulated by MPTP in striatal astrocytes that were labeled with GFAP (red), and combined treatment of β-LAP with MPTP increased the HO-1 expression level. The white arrows indicate the cells that are positive for both GFAP and HO-1. (D) The quantification data showing the proportion of GFAP/HO-1-positive cells to GFAP only-positive cells. * p
Figure Legend Snippet: β-LAP upregulated the expression of HO-1 in striatal astrocytes. (A) Quantitative real-time PCR data showing the mRNA expression of HO-1 in striatum of MPTP-injected mice (n=5 per group). (B) Western blot analysis showing the expression of HO-1 protein in the striatum of MPTP-injected mice (n=5 per group). The quantification data are provided in the right panel. (C) The double IF staining results indicate that the HO-1 protein (green) was upregulated by MPTP in striatal astrocytes that were labeled with GFAP (red), and combined treatment of β-LAP with MPTP increased the HO-1 expression level. The white arrows indicate the cells that are positive for both GFAP and HO-1. (D) The quantification data showing the proportion of GFAP/HO-1-positive cells to GFAP only-positive cells. * p

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

30) Product Images from "Blocking of JB6 Cell Transformation by Tanshinone IIA: Epigenetic Reactivation of Nrf2 Antioxidative Stress Pathway"

Article Title: Blocking of JB6 Cell Transformation by Tanshinone IIA: Epigenetic Reactivation of Nrf2 Antioxidative Stress Pathway

Journal: The AAPS Journal

doi: 10.1208/s12248-014-9666-8

Effect of TIIA on Nrf2 mRNA and protein expression of Nrf2 target genes (HO-1, NQO1, and UGT1A1) in JB6 P+ cells. Cells were incubated with various concentrations of TIIA (2–10 μM) for 5 days. a TIIA increased the mRNA levels of Nrf2 and its downstream representative enzymes including HO-1, NQO1, and UGT1A1; b Western blot images of Nrf2 and its downstream genes including HO-1, NQO1, and UGT1A1; c TIIA (2–10 μM) increased the protein expression of Nrf2 and Nrf2 downstream enzymes. The graphical data are presented as the mean ± SD from three independent experiments. * P
Figure Legend Snippet: Effect of TIIA on Nrf2 mRNA and protein expression of Nrf2 target genes (HO-1, NQO1, and UGT1A1) in JB6 P+ cells. Cells were incubated with various concentrations of TIIA (2–10 μM) for 5 days. a TIIA increased the mRNA levels of Nrf2 and its downstream representative enzymes including HO-1, NQO1, and UGT1A1; b Western blot images of Nrf2 and its downstream genes including HO-1, NQO1, and UGT1A1; c TIIA (2–10 μM) increased the protein expression of Nrf2 and Nrf2 downstream enzymes. The graphical data are presented as the mean ± SD from three independent experiments. * P

Techniques Used: Expressing, Incubation, Western Blot

31) Product Images from "20C, a bibenzyl compound isolated from Gastrodia elata, protects PC12 cells against rotenone-induced apoptosis via activation of the Nrf2/ARE/HO-1 signaling pathway"

Article Title: 20C, a bibenzyl compound isolated from Gastrodia elata, protects PC12 cells against rotenone-induced apoptosis via activation of the Nrf2/ARE/HO-1 signaling pathway

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2015.154

Effects of 20C on the Nrf2/ARE/HO-1 signaling pathway. (A) Western blotting analysis of the levels of the Nrf2 protein in the cytoplasm and nucleus of PC12 cells exposed to rotenone in the presence or absence of various concentrations of 20C. (B) Quantitative analysis of the density of Nrf2 in the cytoplasm and nucleus. (C) Confocal microscopy images of immunofluorescence with an anti-Nrf2 antibody showed the nuclear translocation of Nrf2 in the 20C-treated PC12 cells. The scale bar represents 10 μm. (D, E) Western blotting analysis of the levels of the HO-1 and NQO1 proteins in PC12 cells exposed to rotenone in the presence or absence of 20C. (F, G) RT-PCR analysis of the mRNA levels of HO-1 and NQO1 in PC12 cells exposed to rotenone in the presence or absence of 20C. # P
Figure Legend Snippet: Effects of 20C on the Nrf2/ARE/HO-1 signaling pathway. (A) Western blotting analysis of the levels of the Nrf2 protein in the cytoplasm and nucleus of PC12 cells exposed to rotenone in the presence or absence of various concentrations of 20C. (B) Quantitative analysis of the density of Nrf2 in the cytoplasm and nucleus. (C) Confocal microscopy images of immunofluorescence with an anti-Nrf2 antibody showed the nuclear translocation of Nrf2 in the 20C-treated PC12 cells. The scale bar represents 10 μm. (D, E) Western blotting analysis of the levels of the HO-1 and NQO1 proteins in PC12 cells exposed to rotenone in the presence or absence of 20C. (F, G) RT-PCR analysis of the mRNA levels of HO-1 and NQO1 in PC12 cells exposed to rotenone in the presence or absence of 20C. # P

Techniques Used: Western Blot, Confocal Microscopy, Immunofluorescence, Translocation Assay, Reverse Transcription Polymerase Chain Reaction

32) Product Images from "Protective Actions of Anserine Under Diabetic Conditions"

Article Title: Protective Actions of Anserine Under Diabetic Conditions

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19092751

Effect of co-incubation with high glucose and anserine in human tubular cells (HK-2) on cellular heat shock protein 70 (Hsp70), hemeoxygenase (HO-1), Sirtuin-1 (Sirt-1) and Thioredoxin (Trx). Hsp70 ( A ), HO-1 ( B ), Sirt-1 ( C ) and Trx ( D ) cellular protein concentrations significantly increased in HK-2 cells with glucose stress (25 mM for 24 h), determined by Western blotting, compared to cells incubated with medium containing normal glucose concentration (11 mM). Densitometric units (D.U.) after normalization against β-actin are given ( n = 3). Co-incubation with anserine (0.1 and 1 mM) further increased Hsp70 and HO-1 protein but had no additional effect on Sirt-1 and Trx. Anserine alone does not alter tubular cell defense systems. p
Figure Legend Snippet: Effect of co-incubation with high glucose and anserine in human tubular cells (HK-2) on cellular heat shock protein 70 (Hsp70), hemeoxygenase (HO-1), Sirtuin-1 (Sirt-1) and Thioredoxin (Trx). Hsp70 ( A ), HO-1 ( B ), Sirt-1 ( C ) and Trx ( D ) cellular protein concentrations significantly increased in HK-2 cells with glucose stress (25 mM for 24 h), determined by Western blotting, compared to cells incubated with medium containing normal glucose concentration (11 mM). Densitometric units (D.U.) after normalization against β-actin are given ( n = 3). Co-incubation with anserine (0.1 and 1 mM) further increased Hsp70 and HO-1 protein but had no additional effect on Sirt-1 and Trx. Anserine alone does not alter tubular cell defense systems. p

Techniques Used: Incubation, Western Blot, Concentration Assay

33) Product Images from "Dynorphin activation of kappa opioid receptor protects against epilepsy and seizure-induced brain injury via PI3K/Akt/Nrf2/HO-1 pathway"

Article Title: Dynorphin activation of kappa opioid receptor protects against epilepsy and seizure-induced brain injury via PI3K/Akt/Nrf2/HO-1 pathway

Journal: Cell Cycle

doi: 10.1080/15384101.2018.1562286

Decreased PDYN expression and suppressed PI3K/Akt/Nrf2/HO-1 pathway in epileptiform hippocampal neurons MDA content and SOD activity (a) using commercial kits, cell apoptosis (b) using flow cytometry, mRNA expression of DYN and HO-1 (c) using RT-qPCR, and protein expression of PDYN, total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (d-e) using western blot, in cultured hippocampal neurons exposed to Mg 2+ -free solution or not. *pÂÂ
Figure Legend Snippet: Decreased PDYN expression and suppressed PI3K/Akt/Nrf2/HO-1 pathway in epileptiform hippocampal neurons MDA content and SOD activity (a) using commercial kits, cell apoptosis (b) using flow cytometry, mRNA expression of DYN and HO-1 (c) using RT-qPCR, and protein expression of PDYN, total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (d-e) using western blot, in cultured hippocampal neurons exposed to Mg 2+ -free solution or not. *pÂÂ

Techniques Used: Expressing, Multiple Displacement Amplification, Activity Assay, Flow Cytometry, Cytometry, Quantitative RT-PCR, Western Blot, Cell Culture

Decreased PDYN expression and suppressed PI3K/Akt/Nrf2/HO-1 pathway in a rat model of epilepsy Serum levels of TNF-α, IL-2, and IL-6 (a) using ELISA, MDA content and SOD activity (b) using commercial kits in control and epileptic model rats. In situ cell apoptosis (c) using TUNEL staining (Scale bar: 25.0 μm in overview; 2 μm in CA1 and CA3), mRNA expression of PDYN and HO-1 (d) using RT-qPCR, and protein expression of PDYN, total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (e) using western blot, in the hippocampus of control and epileptic model rats. n = 8/group. *pÂÂ
Figure Legend Snippet: Decreased PDYN expression and suppressed PI3K/Akt/Nrf2/HO-1 pathway in a rat model of epilepsy Serum levels of TNF-α, IL-2, and IL-6 (a) using ELISA, MDA content and SOD activity (b) using commercial kits in control and epileptic model rats. In situ cell apoptosis (c) using TUNEL staining (Scale bar: 25.0 μm in overview; 2 μm in CA1 and CA3), mRNA expression of PDYN and HO-1 (d) using RT-qPCR, and protein expression of PDYN, total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (e) using western blot, in the hippocampus of control and epileptic model rats. n = 8/group. *pÂÂ

Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification, Activity Assay, In Situ, TUNEL Assay, Staining, Quantitative RT-PCR, Western Blot

Dynorphin activation of KOR alleviated seizure-like neuron injury via activation of PI3K/Akt/Nrf2/HO-1 pathway MDA content and SOD activity (a) using commercial kits, cell apoptosis (b) using flow cytometry, mRNA expression of HO-1 (c) using RT-qPCR, and protein expression of total Nrf2, nuclear Nrf2 and HO-1 (d) using western blot, in cultured hippocampal neurons exposed to Mg 2+ -free solution, followed by treatment with dynorphin-A and LY294002 (an inhibitor of PI3K/Akt pathway)/ZnPPIX (an HO-1 inhibitor), both alone or in combination. Cultured hippocampal neurons without any treatment served as the control group. DMSO served as the vehicle of LY294002 and ZnPPIX. Dyn-A, dynorphin-A. LY, LY294002. *pÂÂ
Figure Legend Snippet: Dynorphin activation of KOR alleviated seizure-like neuron injury via activation of PI3K/Akt/Nrf2/HO-1 pathway MDA content and SOD activity (a) using commercial kits, cell apoptosis (b) using flow cytometry, mRNA expression of HO-1 (c) using RT-qPCR, and protein expression of total Nrf2, nuclear Nrf2 and HO-1 (d) using western blot, in cultured hippocampal neurons exposed to Mg 2+ -free solution, followed by treatment with dynorphin-A and LY294002 (an inhibitor of PI3K/Akt pathway)/ZnPPIX (an HO-1 inhibitor), both alone or in combination. Cultured hippocampal neurons without any treatment served as the control group. DMSO served as the vehicle of LY294002 and ZnPPIX. Dyn-A, dynorphin-A. LY, LY294002. *pÂÂ

Techniques Used: Activation Assay, Multiple Displacement Amplification, Activity Assay, Flow Cytometry, Cytometry, Expressing, Quantitative RT-PCR, Western Blot, Cell Culture

Dynorphin activation of KOR activated PI3K/Akt/Nrf2/HO-1 pathway and alleviated seizure-like neuron injury MDA content and SOD activity (a) using commercial kits, cell apoptosis (b) using flow cytometry, mRNA expression of HO-1 (c) using RT-qPCR, and protein expression of total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (d) using western blot, in cultured hippocampal neurons exposed to Mg 2+ -free solution, followed by treatment with the KOR agonist dynorphin-A and the KOR antagonist GNTI, both alone or in combination. Cultured hippocampal neurons without any treatment served as the control group. Dyn-A, dynorphin-A. *pÂÂ
Figure Legend Snippet: Dynorphin activation of KOR activated PI3K/Akt/Nrf2/HO-1 pathway and alleviated seizure-like neuron injury MDA content and SOD activity (a) using commercial kits, cell apoptosis (b) using flow cytometry, mRNA expression of HO-1 (c) using RT-qPCR, and protein expression of total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (d) using western blot, in cultured hippocampal neurons exposed to Mg 2+ -free solution, followed by treatment with the KOR agonist dynorphin-A and the KOR antagonist GNTI, both alone or in combination. Cultured hippocampal neurons without any treatment served as the control group. Dyn-A, dynorphin-A. *pÂÂ

Techniques Used: Activation Assay, Multiple Displacement Amplification, Activity Assay, Flow Cytometry, Cytometry, Expressing, Quantitative RT-PCR, Western Blot, Cell Culture

PDYN overexpression alleviated pilocarpine-induced epilepsy and neuronal apoptosis in rats Serum levels of TNF-α, IL-2, and IL-6 (a) using ELISA, MDA content and SOD activity (b) using commercial kits, in the groups of model, LV-NC, and LV-Dyn. In situ cell apoptosis (c) using TUNEL staining (scale bar: 25.0 μm in overview; 2 μm in CA1 and CA3), mRNA expression of PDYN (d) and HO-1 (e) using RT-qPCR, and protein expression of PDYN, total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (f) using western blot, in the rat hippocampus in the groups of model, LV-NC, and LV-PDYN. n = 8/group. *pÂÂ
Figure Legend Snippet: PDYN overexpression alleviated pilocarpine-induced epilepsy and neuronal apoptosis in rats Serum levels of TNF-α, IL-2, and IL-6 (a) using ELISA, MDA content and SOD activity (b) using commercial kits, in the groups of model, LV-NC, and LV-Dyn. In situ cell apoptosis (c) using TUNEL staining (scale bar: 25.0 μm in overview; 2 μm in CA1 and CA3), mRNA expression of PDYN (d) and HO-1 (e) using RT-qPCR, and protein expression of PDYN, total Nrf2, nuclear Nrf2, HO-1, PI3K, as well as phosphorylation level of Akt (p-Akt) (f) using western blot, in the rat hippocampus in the groups of model, LV-NC, and LV-PDYN. n = 8/group. *pÂÂ

Techniques Used: Over Expression, Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification, Activity Assay, In Situ, TUNEL Assay, Staining, Expressing, Quantitative RT-PCR, Western Blot

34) Product Images from "N-Octanoyl Dopamine Inhibits the Expression of a Subset of ?B Regulated Genes: Potential Role of p65 Ser276 Phosphorylation"

Article Title: N-Octanoyl Dopamine Inhibits the Expression of a Subset of ?B Regulated Genes: Potential Role of p65 Ser276 Phosphorylation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0073122

Partial silencing of Nrf-2 and HO-1 expression does not abrogate NOD mediated inhibition of VCAM-1. HUVECs were transfected with control siRNA, HO-1 siRNA or Nrf-2 siRNA. One day after transfection the cells were stimulated for 24 hrs with TNF-α alone (10 ng/ml), NOD alone (50 µM) or in combination of both. Cells that were not stimulated were included in each experiment. The expression of VCAM-1, Nrf-2 and HO-1 was assessed by western blotting. GAPDH was used as loading control. The results of a representative experiment are shown. A total of 3 independent experiments with different HUVEC cultures were performed.
Figure Legend Snippet: Partial silencing of Nrf-2 and HO-1 expression does not abrogate NOD mediated inhibition of VCAM-1. HUVECs were transfected with control siRNA, HO-1 siRNA or Nrf-2 siRNA. One day after transfection the cells were stimulated for 24 hrs with TNF-α alone (10 ng/ml), NOD alone (50 µM) or in combination of both. Cells that were not stimulated were included in each experiment. The expression of VCAM-1, Nrf-2 and HO-1 was assessed by western blotting. GAPDH was used as loading control. The results of a representative experiment are shown. A total of 3 independent experiments with different HUVEC cultures were performed.

Techniques Used: Expressing, Inhibition, Transfection, Western Blot

HO-1 induction by NOD is not required for inhibition of NFκB. (A) HUVECs were pre-treated for 2 hrs with 5 µg/ml of cyclohexamide (+ CyHx) or left untreated (− CyHx). Hereafter, cells were stimulated for 8 hrs with 10 ng/ml of TNF-α in the presence (+) or absence (−) of 50 µM of NOD. In case the cells were pre-treated with CyHx, this was present during the whole period of stimulation. In case cells were not treated with CyHx, this was absent during stimulation. Western blotting of the cytoplasmic fractions revealed that de novo protein synthesis was effectively inhibited by CyHx. Note that VCAM-1 is not induced by TNF-α in the presence of CyHx. Also in the combination of TNF-α+NOD the induction of HO-1 did not occur. GAPDH was used as loading control. (B) Nuclear extracts were prepared and assessed for NFκB activation by means of EMSA. Specificity of the bands was assessed by adding an excess of unlabelled NFκB consensus (cold consensus (CC)) or mutated (cold mutated (CM)) oligonucleotides to the samples. To demonstrate the presence of p50 and p65 in the shifted bands super-shifts (SS) were performed by adding anti-p50 or anti-p65 monoclonal antibodies to the samples. In A und B the results of a representative experiment are shown. A total of 4 independent experiments with different HUVEC cultures were performed.
Figure Legend Snippet: HO-1 induction by NOD is not required for inhibition of NFκB. (A) HUVECs were pre-treated for 2 hrs with 5 µg/ml of cyclohexamide (+ CyHx) or left untreated (− CyHx). Hereafter, cells were stimulated for 8 hrs with 10 ng/ml of TNF-α in the presence (+) or absence (−) of 50 µM of NOD. In case the cells were pre-treated with CyHx, this was present during the whole period of stimulation. In case cells were not treated with CyHx, this was absent during stimulation. Western blotting of the cytoplasmic fractions revealed that de novo protein synthesis was effectively inhibited by CyHx. Note that VCAM-1 is not induced by TNF-α in the presence of CyHx. Also in the combination of TNF-α+NOD the induction of HO-1 did not occur. GAPDH was used as loading control. (B) Nuclear extracts were prepared and assessed for NFκB activation by means of EMSA. Specificity of the bands was assessed by adding an excess of unlabelled NFκB consensus (cold consensus (CC)) or mutated (cold mutated (CM)) oligonucleotides to the samples. To demonstrate the presence of p50 and p65 in the shifted bands super-shifts (SS) were performed by adding anti-p50 or anti-p65 monoclonal antibodies to the samples. In A und B the results of a representative experiment are shown. A total of 4 independent experiments with different HUVEC cultures were performed.

Techniques Used: Inhibition, Western Blot, Activation Assay

Influence of NOD on the expression of adhesion molecules and HO-1. (A) HUVECs were stimulated for 24 hrs with 10 ng/ml of TNF-α in the presence of different concentrations of NOD. The expression of VCAM-1 and HO-1 was assessed by western blotting. GAPDH was used as loading control. (B) HUVECs were stimulated as described in A. The expression of ICAM-1 and E-selectin was assessed by FACS analysis. (C) To demonstrate that the induction of HO-1 by NOD was independent of TNF-α, HUVECs were stimulated for 24 hrs with 10 ng/ml of TNF-α alone, 100 µM of NOD alone or in combination of both. HUVECs cultured in medium served as control. The expression of VCAM-1 and HO-1 was assessed by western blotting. The results shown in A, B and C are representative experiments. A total of 6 independent experiments with different HUVEC cultures were performed. All westernblots have been scanned and statistics was performed on the ratio of the optical density of protein of interest/optical density of GAPDH. Significant inhibition of VCAM-1 expression occurred at concentrations above 12.5 µM and HO-1 induction already at 1 µM ( p
Figure Legend Snippet: Influence of NOD on the expression of adhesion molecules and HO-1. (A) HUVECs were stimulated for 24 hrs with 10 ng/ml of TNF-α in the presence of different concentrations of NOD. The expression of VCAM-1 and HO-1 was assessed by western blotting. GAPDH was used as loading control. (B) HUVECs were stimulated as described in A. The expression of ICAM-1 and E-selectin was assessed by FACS analysis. (C) To demonstrate that the induction of HO-1 by NOD was independent of TNF-α, HUVECs were stimulated for 24 hrs with 10 ng/ml of TNF-α alone, 100 µM of NOD alone or in combination of both. HUVECs cultured in medium served as control. The expression of VCAM-1 and HO-1 was assessed by western blotting. The results shown in A, B and C are representative experiments. A total of 6 independent experiments with different HUVEC cultures were performed. All westernblots have been scanned and statistics was performed on the ratio of the optical density of protein of interest/optical density of GAPDH. Significant inhibition of VCAM-1 expression occurred at concentrations above 12.5 µM and HO-1 induction already at 1 µM ( p

Techniques Used: Expressing, Western Blot, FACS, Cell Culture, Inhibition

A redox moiety and sufficient hydrophobicity are required for NOD mediated inhibition of VCAM-1 expression and for induction of HO-1. HUVECs were stimulated with 10 ng/ml of TNF-α alone (−) or stimulated with TNF-α in the presence of the different compounds shown in figure 6 . Cells that were left untreated (medium) served as control. The expression of VCAM-1 and HO-1 was assessed by western blotting. GAPDH was used as loading control. The results of a representative experiment are shown. A total of 3 independent experiments with different HUVEC cultures were performed.
Figure Legend Snippet: A redox moiety and sufficient hydrophobicity are required for NOD mediated inhibition of VCAM-1 expression and for induction of HO-1. HUVECs were stimulated with 10 ng/ml of TNF-α alone (−) or stimulated with TNF-α in the presence of the different compounds shown in figure 6 . Cells that were left untreated (medium) served as control. The expression of VCAM-1 and HO-1 was assessed by western blotting. GAPDH was used as loading control. The results of a representative experiment are shown. A total of 3 independent experiments with different HUVEC cultures were performed.

Techniques Used: Inhibition, Expressing, Western Blot

35) Product Images from "Astragalus membranaceus Extract Attenuates Inflammation and Oxidative Stress in Intestinal Epithelial Cells via NF-κB Activation and Nrf2 Response"

Article Title: Astragalus membranaceus Extract Attenuates Inflammation and Oxidative Stress in Intestinal Epithelial Cells via NF-κB Activation and Nrf2 Response

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19030800

( A ) Effect of Astragalus membranaceus extract (50 μg/mL) alone and in the presence of H 2 O 2 on the translocation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) into the cellular nuclei, evaluated by immunofluorescence analysis. The blue fluorescence identified the nuclei while the green fluorescence indicated Nrf2. ( B ) Effect of Astragalus membranaceus extract (5–100 μg/mL) in the presence of H 2 O 2 on heme oxygenase 1 (HO-1) expression, evaluated by the cytofluorimetric technique ( C ) and on NAD(P)H quinone dehydrogenase 1 (NQO1) in IEC-6 cells, evaluated by the cytofluorimetric technique. The dark grey bars (B, C) represent IEC-6 cells treated with H 2 O 2 alone. Data are expressed as mean ± SEM.°°° denotes p
Figure Legend Snippet: ( A ) Effect of Astragalus membranaceus extract (50 μg/mL) alone and in the presence of H 2 O 2 on the translocation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) into the cellular nuclei, evaluated by immunofluorescence analysis. The blue fluorescence identified the nuclei while the green fluorescence indicated Nrf2. ( B ) Effect of Astragalus membranaceus extract (5–100 μg/mL) in the presence of H 2 O 2 on heme oxygenase 1 (HO-1) expression, evaluated by the cytofluorimetric technique ( C ) and on NAD(P)H quinone dehydrogenase 1 (NQO1) in IEC-6 cells, evaluated by the cytofluorimetric technique. The dark grey bars (B, C) represent IEC-6 cells treated with H 2 O 2 alone. Data are expressed as mean ± SEM.°°° denotes p

Techniques Used: Translocation Assay, Derivative Assay, Immunofluorescence, Fluorescence, Expressing

36) Product Images from "Phloretin Attenuates Allergic Airway Inflammation and Oxidative Stress in Asthmatic Mice"

Article Title: Phloretin Attenuates Allergic Airway Inflammation and Oxidative Stress in Asthmatic Mice

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00134

Phloretin (PT) effects on oxidative stress factors . (A) Western blot shows PT modulation of HO-1 and Nrf2 expression in lung tissue of normal (N) and OVA-stimulated (OVA) mice, without or with PT (PT5-20) treatment. (B) Malondialdehyde (MDA) activity and (C) GSH activity in lung tissues of mice. Data are presented as the mean ± SEM. * p
Figure Legend Snippet: Phloretin (PT) effects on oxidative stress factors . (A) Western blot shows PT modulation of HO-1 and Nrf2 expression in lung tissue of normal (N) and OVA-stimulated (OVA) mice, without or with PT (PT5-20) treatment. (B) Malondialdehyde (MDA) activity and (C) GSH activity in lung tissues of mice. Data are presented as the mean ± SEM. * p

Techniques Used: Western Blot, Expressing, Mouse Assay, Multiple Displacement Amplification, Activity Assay

37) Product Images from "Hemopexin alleviates cognitive dysfunction after focal cerebral ischemia-reperfusion injury in rats"

Article Title: Hemopexin alleviates cognitive dysfunction after focal cerebral ischemia-reperfusion injury in rats

Journal: BMC Anesthesiology

doi: 10.1186/s12871-019-0681-2

HPX up-regulated the expression of HO-1 on day 7 after focal cerebral I/R injury. When compared with the rats in the sham group, those with focal cerebral I/R injury exhibited an increase in the HO-1 protein expression on day 7 after I/R injury. The HO-1 protein expression was increased in the vehicle and MCAO group. HPX injection up-regulated the HO-1 expression. * P
Figure Legend Snippet: HPX up-regulated the expression of HO-1 on day 7 after focal cerebral I/R injury. When compared with the rats in the sham group, those with focal cerebral I/R injury exhibited an increase in the HO-1 protein expression on day 7 after I/R injury. The HO-1 protein expression was increased in the vehicle and MCAO group. HPX injection up-regulated the HO-1 expression. * P

Techniques Used: Expressing, Injection

Intracerebroventricular HPX injection enhanced the neovascularization of the ischemic penumbra hippocampus in rats after focal cerebral I/R injury. Compared with the respective densities in the sham group, the new vessel densities in the ischemic penumbra hippocampus ( a ) in rats of the MCAO group were higher at 7 days after focal cerebral I/R injury. Compared with the respective densities in the vehicle group, the new vessel densities of the ischemic penumbra hippocampus ( a ) in rats of the HPX group were markedly increased at 7 days after focal cerebral I/R injury. When the inhibitor of HO-1, ZnPPIX, was given along with HPX to rats after focal cerebral I/R injury, the positive effect of HPX on enhancing the neovascularization of the ischemic penumbra hippocampus ( a , b ) was significantly blocked. However, there was no significant difference in the new vessel density of the ischemic hippocampus in rats in the MCAO and vehicle group at 7 days after focal cerebral I/R injury ( a , b ). * P
Figure Legend Snippet: Intracerebroventricular HPX injection enhanced the neovascularization of the ischemic penumbra hippocampus in rats after focal cerebral I/R injury. Compared with the respective densities in the sham group, the new vessel densities in the ischemic penumbra hippocampus ( a ) in rats of the MCAO group were higher at 7 days after focal cerebral I/R injury. Compared with the respective densities in the vehicle group, the new vessel densities of the ischemic penumbra hippocampus ( a ) in rats of the HPX group were markedly increased at 7 days after focal cerebral I/R injury. When the inhibitor of HO-1, ZnPPIX, was given along with HPX to rats after focal cerebral I/R injury, the positive effect of HPX on enhancing the neovascularization of the ischemic penumbra hippocampus ( a , b ) was significantly blocked. However, there was no significant difference in the new vessel density of the ischemic hippocampus in rats in the MCAO and vehicle group at 7 days after focal cerebral I/R injury ( a , b ). * P

Techniques Used: Injection

Intracerebroventricular HPX injection improved the long-term spatial learning and memory ability in rats after focal I/R injury. The baseline escape latencies in the spatial probe test among the five groups before sham operation and focal cerebral I/R injury (− 24 h) were not significantly different ( a ). Compared with rats in the sham group, rats in the MCAO group needed more time to find the platform hidden under water in the spatial probe test ( b ) at 2–7 d after focal I/R injury, spent less time in the target quadrant ( c ) and crossed the platform fewer times in the hidden platform test ( d ) at 7 d after focal I/R. Compared with the rats in the vehicle group, the rats in the HPX group exhibited a shorter escape latency on day 2 to 7 after focal cerebral I/R injury in the spatial probe test ( b ) and a increase in the time spent in the target quadrant ( c ) and number of platform crossings ( d ) in the hidden platform test at 7 d after focal I/R. The inhibitor of HO-1, ZnPPIX, blocked the effect of HPX on shortening the escape latency in the spatial probe test and increasing the time spent in the target quadrant ( c ) and number of platform crossings ( d ) in the hidden platform test. However, there were no significant differences in the escape latency in the spatial probe test ( b ), the time spent in the target quadrant ( c ) or the number of platform crossings ( d ) in the hidden platform test between the MCAO and vehicle group after focal cerebral I/R injury. * P
Figure Legend Snippet: Intracerebroventricular HPX injection improved the long-term spatial learning and memory ability in rats after focal I/R injury. The baseline escape latencies in the spatial probe test among the five groups before sham operation and focal cerebral I/R injury (− 24 h) were not significantly different ( a ). Compared with rats in the sham group, rats in the MCAO group needed more time to find the platform hidden under water in the spatial probe test ( b ) at 2–7 d after focal I/R injury, spent less time in the target quadrant ( c ) and crossed the platform fewer times in the hidden platform test ( d ) at 7 d after focal I/R. Compared with the rats in the vehicle group, the rats in the HPX group exhibited a shorter escape latency on day 2 to 7 after focal cerebral I/R injury in the spatial probe test ( b ) and a increase in the time spent in the target quadrant ( c ) and number of platform crossings ( d ) in the hidden platform test at 7 d after focal I/R. The inhibitor of HO-1, ZnPPIX, blocked the effect of HPX on shortening the escape latency in the spatial probe test and increasing the time spent in the target quadrant ( c ) and number of platform crossings ( d ) in the hidden platform test. However, there were no significant differences in the escape latency in the spatial probe test ( b ), the time spent in the target quadrant ( c ) or the number of platform crossings ( d ) in the hidden platform test between the MCAO and vehicle group after focal cerebral I/R injury. * P

Techniques Used: Injection

38) Product Images from "KMS99220 Exerts Anti-Inflammatory Effects, Activates the Nrf2 Signaling and Interferes with IKK, JNK and p38 MAPK via HO-1"

Article Title: KMS99220 Exerts Anti-Inflammatory Effects, Activates the Nrf2 Signaling and Interferes with IKK, JNK and p38 MAPK via HO-1

Journal: Molecules and Cells

doi: 10.14348/molcells.2019.0129

KMS99220 activates the Nrf2 signaling pathway and induces HO-1 production in microglia (A–I) BV-2 cells were treated with various concentrations of KMS99220. (A) Nuclear extracts from BV-2 cells were obtained after incubation with KMS99220 for 3 h, and the amount of nuclear Nrf2 (57 kDa) was analyzed by Western blot analysis, using lamin B (67 kDa) as an internal control. After incubation with KMS99220 for 24 h, Western blot analysis was performed for HO-1 (32 kDa) (B), NQO1 (31 kDa) (C), GCLC (73 kDa) (D), and GCLM (31 kDa) (E) using β-actin as an internal control. After incubation for 6 h, RT-PCR was performed for HO-1 (F), NQO1 (G), GCLC (H), and GCLM (I) using GAPDH as an internal control. The data are expressed as fold changes compared with untreated control ± SEM (n = 3); # P
Figure Legend Snippet: KMS99220 activates the Nrf2 signaling pathway and induces HO-1 production in microglia (A–I) BV-2 cells were treated with various concentrations of KMS99220. (A) Nuclear extracts from BV-2 cells were obtained after incubation with KMS99220 for 3 h, and the amount of nuclear Nrf2 (57 kDa) was analyzed by Western blot analysis, using lamin B (67 kDa) as an internal control. After incubation with KMS99220 for 24 h, Western blot analysis was performed for HO-1 (32 kDa) (B), NQO1 (31 kDa) (C), GCLC (73 kDa) (D), and GCLM (31 kDa) (E) using β-actin as an internal control. After incubation for 6 h, RT-PCR was performed for HO-1 (F), NQO1 (G), GCLC (H), and GCLM (I) using GAPDH as an internal control. The data are expressed as fold changes compared with untreated control ± SEM (n = 3); # P

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

HO-1 mediates the KMS99220-induced inhibition of IKK, p38 and JNK (A and B) BV-2 cells were transfected with control or HO-1 siRNA for 24 h. (A) Western blot results of HO-1. (B) Western blot results of total and phosphorylated levels of IKK, JNK, p38 MAPK, and ERK in BV-2 cells after treatment with 10 μM KMS99220 for 1 h and incubation with 0.2 μg/ml LPS for 0.5 h. (C) Western blot results of total and phosphorylated levels of IKK, JNK, p38 MAPK, and ERK after pretreatment with SnPP for 1 h and subsequent treatment with 10 μM KMS99220 for 1 h followed by 0.2 μg/ml LPS for 0.5 h.
Figure Legend Snippet: HO-1 mediates the KMS99220-induced inhibition of IKK, p38 and JNK (A and B) BV-2 cells were transfected with control or HO-1 siRNA for 24 h. (A) Western blot results of HO-1. (B) Western blot results of total and phosphorylated levels of IKK, JNK, p38 MAPK, and ERK in BV-2 cells after treatment with 10 μM KMS99220 for 1 h and incubation with 0.2 μg/ml LPS for 0.5 h. (C) Western blot results of total and phosphorylated levels of IKK, JNK, p38 MAPK, and ERK after pretreatment with SnPP for 1 h and subsequent treatment with 10 μM KMS99220 for 1 h followed by 0.2 μg/ml LPS for 0.5 h.

Techniques Used: Inhibition, Transfection, Western Blot, Incubation

39) Product Images from "20C, a bibenzyl compound isolated from Gastrodia elata, protects PC12 cells against rotenone-induced apoptosis via activation of the Nrf2/ARE/HO-1 signaling pathway"

Article Title: 20C, a bibenzyl compound isolated from Gastrodia elata, protects PC12 cells against rotenone-induced apoptosis via activation of the Nrf2/ARE/HO-1 signaling pathway

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2015.154

Effects of 20C on the Nrf2/ARE/HO-1 signaling pathway. (A) Western blotting analysis of the levels of the Nrf2 protein in the cytoplasm and nucleus of PC12 cells exposed to rotenone in the presence or absence of various concentrations of 20C. (B) Quantitative
Figure Legend Snippet: Effects of 20C on the Nrf2/ARE/HO-1 signaling pathway. (A) Western blotting analysis of the levels of the Nrf2 protein in the cytoplasm and nucleus of PC12 cells exposed to rotenone in the presence or absence of various concentrations of 20C. (B) Quantitative

Techniques Used: Western Blot

40) Product Images from "Antioxidant Properties of Buffalo-Milk Dairy Products: A β-Lg Peptide Released after Gastrointestinal Digestion of Buffalo Ricotta Cheese Reduces Oxidative Stress in Intestinal Epithelial Cells"

Article Title: Antioxidant Properties of Buffalo-Milk Dairy Products: A β-Lg Peptide Released after Gastrointestinal Digestion of Buffalo Ricotta Cheese Reduces Oxidative Stress in Intestinal Epithelial Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19071955

( a ) Effect on HO-1, ( b ) NQO1, and ( c ) SOD expression of BRP in the IEC-6 cells, evaluated by cytofluorimetric technique. Values, mean ± s.e.m., are expressed as the % of inhibition of HO-1, NQO1, and SOD expression vs. H 2 O 2 alone-treated cells. *** and * denote p
Figure Legend Snippet: ( a ) Effect on HO-1, ( b ) NQO1, and ( c ) SOD expression of BRP in the IEC-6 cells, evaluated by cytofluorimetric technique. Values, mean ± s.e.m., are expressed as the % of inhibition of HO-1, NQO1, and SOD expression vs. H 2 O 2 alone-treated cells. *** and * denote p

Techniques Used: Expressing, Inhibition

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Blocking Assay:

Article Title: Association of HMGB1 with oxidative stress markers and regulators in PDR
Article Snippet: .. They were left untreated or were treated with 100 ng/ml of cytokine-HMGB1, lipopolysaccharide free (Cat. No. REHM120, IBL International Corporation) for 24 h and were fixed with ice-cold acetone or 4% paraformaldehyde followed by blocking with 10% goat serum for 1 h. Without being washed, the cells were incubated with anti-HO-1 (1:100, sc-10789, Santa Cruz Biotechnology, Inc.), anti-VAP-1 (1:100, ab196739, Abcam), or anti-8OHdG (1:50, ab62623, Abcam) primary antibodies overnight at 4 °C. .. After washing, cells were incubated with secondary antibodies conjugated with appropriate fluorophores (Alexa Fluor 488 or 555) diluted in 10% goat serum (1:1000) for 1 h at room temperature.

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  • 95
    Santa Cruz Biotechnology anti ho 1
    Digoxin decreased Nrf2 at transcriptional level through inhibiting PI3K/Akt pathway in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on protein levels of p-JNK, JNK, p-ERK1/2, ERK1/2, p-P38 and P38. (C–D) Effects of digoxin on protein levels of PI3K, p-Akt, Akt. (E–F) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2, NQO1, <t>HO-1,</t> and GCLC were detected by Western blot. (G–H) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, Nrf2 mRNA levels were detected by qRT-PCR. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P
    Anti Ho 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology mouse monoclonal anti ho 1 sc 390991 anti ampk
    Effect of curcumin on apoptotic, oxidative stress and lipid oxidative signaling pathway associated proteins in kidney tissue. ( a ) Representative western blots depicting protein levels of PI 3 K, p-PI 3 K Akt, p-Akt, <t>AMPK,</t> p-AMPK, cleaved caspase-3, cleaved caspase-9, Nrf2 and <t>HO-1</t> in kidney tissue. ( b ) The CUR + AKI group showed significantly increased levels of p-PI 3 K, p-Akt, p-AMPK, Nrf2, and HO-1, and decreased levels of cleaved caspase-3, cleaved caspase-9 compared to the AKI and CO + AKI groups. Data are the means ± SD (n = 10). Statistical significance: *p
    Mouse Monoclonal Anti Ho 1 Sc 390991 Anti Ampk, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti ho 1 sc 390991 anti ampk/product/Santa Cruz Biotechnology
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    88
    Santa Cruz Biotechnology ho 1 antibodies
    CAE-induced activation of NRF2/HO1 pathway was not linked with AHR activation. (a, b) HaCaT cells were transfected with control siRNA, AhR siRNA, or NrF2 siRNA using the DharmaFECT® Duo transfection reagent. After 24 h, the cells were incubated with CAE (100 μ g/ml) in the presence of B[a]P for 24 h and then subjected to real-time PCR analysis for <t>HO-1</t> and CYP1A1 . ∗ p
    Ho 1 Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ho 1 antibodies/product/Santa Cruz Biotechnology
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    ho 1 antibodies - by Bioz Stars, 2020-05
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    Digoxin decreased Nrf2 at transcriptional level through inhibiting PI3K/Akt pathway in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on protein levels of p-JNK, JNK, p-ERK1/2, ERK1/2, p-P38 and P38. (C–D) Effects of digoxin on protein levels of PI3K, p-Akt, Akt. (E–F) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2, NQO1, HO-1, and GCLC were detected by Western blot. (G–H) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, Nrf2 mRNA levels were detected by qRT-PCR. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

    Journal: Redox Biology

    Article Title: Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway

    doi: 10.1016/j.redox.2019.101131

    Figure Lengend Snippet: Digoxin decreased Nrf2 at transcriptional level through inhibiting PI3K/Akt pathway in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on protein levels of p-JNK, JNK, p-ERK1/2, ERK1/2, p-P38 and P38. (C–D) Effects of digoxin on protein levels of PI3K, p-Akt, Akt. (E–F) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2, NQO1, HO-1, and GCLC were detected by Western blot. (G–H) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, Nrf2 mRNA levels were detected by qRT-PCR. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

    Article Snippet: Anti-NQO1, anti-HO-1 and anti-GCLC antibodies were obtained from Santa Cruz Biotechnology (Texas, USA).

    Techniques: Western Blot, Quantitative RT-PCR

    Digoxin increased the sensitivity of SW1990/Gem and Panc-1/Gem cells to gemcitabine by inhibiting Nrf2 signaling. (A–B) Effects of Nrf2 knockdown on protein levels of Nrf2, NQO1, HO-1, and GCLC in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of Nrf2 knockdown on reversing drug resistance of gemcitabine in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

    Journal: Redox Biology

    Article Title: Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway

    doi: 10.1016/j.redox.2019.101131

    Figure Lengend Snippet: Digoxin increased the sensitivity of SW1990/Gem and Panc-1/Gem cells to gemcitabine by inhibiting Nrf2 signaling. (A–B) Effects of Nrf2 knockdown on protein levels of Nrf2, NQO1, HO-1, and GCLC in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of Nrf2 knockdown on reversing drug resistance of gemcitabine in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

    Article Snippet: Anti-NQO1, anti-HO-1 and anti-GCLC antibodies were obtained from Santa Cruz Biotechnology (Texas, USA).

    Techniques:

    Nrf2 signaling was upregulated in SW1990/Gem and Panc-1/Gem cells. (A–B) The cytotoxicity of gemcitabine to SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (C–D) Western blot was used to detect the protein level of Nrf2, NQO1, HO-1, and GCLC in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (E–F) The expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

    Journal: Redox Biology

    Article Title: Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway

    doi: 10.1016/j.redox.2019.101131

    Figure Lengend Snippet: Nrf2 signaling was upregulated in SW1990/Gem and Panc-1/Gem cells. (A–B) The cytotoxicity of gemcitabine to SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (C–D) Western blot was used to detect the protein level of Nrf2, NQO1, HO-1, and GCLC in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. (E–F) The expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P

    Article Snippet: Anti-NQO1, anti-HO-1 and anti-GCLC antibodies were obtained from Santa Cruz Biotechnology (Texas, USA).

    Techniques: Western Blot, Expressing

    Digoxin inhibited the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of digoxin on protein levels of NQO1, HO-1 and GCLC in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as **P

    Journal: Redox Biology

    Article Title: Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway

    doi: 10.1016/j.redox.2019.101131

    Figure Lengend Snippet: Digoxin inhibited the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (A–B) Effects of digoxin on the expressions of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. The colors of the heatmap reflect log 2 -expression levels of Nrf2 target genes in SW1990/Gem and Panc-1/Gem cells. (C–D) Effects of digoxin on protein levels of NQO1, HO-1 and GCLC in SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as **P

    Article Snippet: Anti-NQO1, anti-HO-1 and anti-GCLC antibodies were obtained from Santa Cruz Biotechnology (Texas, USA).

    Techniques: Expressing

    Digoxin sensitized SW1990/Gem cells-derived xenografts to gemcitabine treatment by inhibiting Nrf2 signaling. (A–D) Digoxin sensitized SW1990/Gem-shControl cells-derived xenografts to gemcitabine treatment. (E–H) Digoxin could not sensitize SW1990/Gem-shNrf2 cells-derived xenografts to gemcitabine treatment. (I) Effects of digoxin on the protein levels of Nrf2, NQO1, HO-1, and GCLC in tumor tissues. (J) Tumor tissues were subjected to IHC-Nrf2, IHC-NQO1, IHC-HO-1, IHC-GCLC, IHC-Ki67 and TUNEL staining. All images were shown at ×200. Data were expressed as mean ± SD, n = 6. Significant differences were indicated as ***P

    Journal: Redox Biology

    Article Title: Digoxin sensitizes gemcitabine-resistant pancreatic cancer cells to gemcitabine via inhibiting Nrf2 signaling pathway

    doi: 10.1016/j.redox.2019.101131

    Figure Lengend Snippet: Digoxin sensitized SW1990/Gem cells-derived xenografts to gemcitabine treatment by inhibiting Nrf2 signaling. (A–D) Digoxin sensitized SW1990/Gem-shControl cells-derived xenografts to gemcitabine treatment. (E–H) Digoxin could not sensitize SW1990/Gem-shNrf2 cells-derived xenografts to gemcitabine treatment. (I) Effects of digoxin on the protein levels of Nrf2, NQO1, HO-1, and GCLC in tumor tissues. (J) Tumor tissues were subjected to IHC-Nrf2, IHC-NQO1, IHC-HO-1, IHC-GCLC, IHC-Ki67 and TUNEL staining. All images were shown at ×200. Data were expressed as mean ± SD, n = 6. Significant differences were indicated as ***P

    Article Snippet: Anti-NQO1, anti-HO-1 and anti-GCLC antibodies were obtained from Santa Cruz Biotechnology (Texas, USA).

    Techniques: Derivative Assay, Immunohistochemistry, TUNEL Assay, Staining

    Translocation of Nrf2-IQGAP1 complex into the nucleus by calcium treatment and induction of HO-1 protein. (A) To verify the involvement of calcium in endogenous Nrf2-IQGAP1 complexes, HeLa cells were transfected with pEGFP-Nrf2 (8 μg/100 mm

    Journal: Antioxidants & Redox Signaling

    Article Title: Identification and Functional Studies of a New Nrf2 Partner IQGAP1: A Critical Role in the Stability and Transactivation of Nrf2

    doi: 10.1089/ars.2012.4586

    Figure Lengend Snippet: Translocation of Nrf2-IQGAP1 complex into the nucleus by calcium treatment and induction of HO-1 protein. (A) To verify the involvement of calcium in endogenous Nrf2-IQGAP1 complexes, HeLa cells were transfected with pEGFP-Nrf2 (8 μg/100 mm

    Article Snippet: The anti-IQGAP (H-109), anti-GFP (B-2), anti-Dsred (C-20), and anti-Maf F/G/K (H-100), anti-HO-1 (C-20), anti-Bach1 (C-20), anti-actin, anti-Lamin A primary antibodies, and secondary antibodies were purchased from Santa Cruz Biotechnology.

    Techniques: Translocation Assay, Transfection

    IQGAP1 enhances the expression of Nrf2 and induction of HO-1 expression as well as the stability of Nrf2. (A) Based on mass data shown in , the newly identified IQGAP1 was then tested to see whether it could regulate the expression of HO-1, which

    Journal: Antioxidants & Redox Signaling

    Article Title: Identification and Functional Studies of a New Nrf2 Partner IQGAP1: A Critical Role in the Stability and Transactivation of Nrf2

    doi: 10.1089/ars.2012.4586

    Figure Lengend Snippet: IQGAP1 enhances the expression of Nrf2 and induction of HO-1 expression as well as the stability of Nrf2. (A) Based on mass data shown in , the newly identified IQGAP1 was then tested to see whether it could regulate the expression of HO-1, which

    Article Snippet: The anti-IQGAP (H-109), anti-GFP (B-2), anti-Dsred (C-20), and anti-Maf F/G/K (H-100), anti-HO-1 (C-20), anti-Bach1 (C-20), anti-actin, anti-Lamin A primary antibodies, and secondary antibodies were purchased from Santa Cruz Biotechnology.

    Techniques: Expressing

    siIQGAP1 decreases the stability of Nrf2 and attenuates HO-1 expression and ARE-luciferase activity. (A) HeLa cells were transfected with siIQGAP1 for 72 h using Lipofectamin 2000 reagent to silence or knockdown the endogenous IQGAP1 gene expression.

    Journal: Antioxidants & Redox Signaling

    Article Title: Identification and Functional Studies of a New Nrf2 Partner IQGAP1: A Critical Role in the Stability and Transactivation of Nrf2

    doi: 10.1089/ars.2012.4586

    Figure Lengend Snippet: siIQGAP1 decreases the stability of Nrf2 and attenuates HO-1 expression and ARE-luciferase activity. (A) HeLa cells were transfected with siIQGAP1 for 72 h using Lipofectamin 2000 reagent to silence or knockdown the endogenous IQGAP1 gene expression.

    Article Snippet: The anti-IQGAP (H-109), anti-GFP (B-2), anti-Dsred (C-20), and anti-Maf F/G/K (H-100), anti-HO-1 (C-20), anti-Bach1 (C-20), anti-actin, anti-Lamin A primary antibodies, and secondary antibodies were purchased from Santa Cruz Biotechnology.

    Techniques: Expressing, Luciferase, Activity Assay, Transfection

    Effect of curcumin on apoptotic, oxidative stress and lipid oxidative signaling pathway associated proteins in kidney tissue. ( a ) Representative western blots depicting protein levels of PI 3 K, p-PI 3 K Akt, p-Akt, AMPK, p-AMPK, cleaved caspase-3, cleaved caspase-9, Nrf2 and HO-1 in kidney tissue. ( b ) The CUR + AKI group showed significantly increased levels of p-PI 3 K, p-Akt, p-AMPK, Nrf2, and HO-1, and decreased levels of cleaved caspase-3, cleaved caspase-9 compared to the AKI and CO + AKI groups. Data are the means ± SD (n = 10). Statistical significance: *p

    Journal: Scientific Reports

    Article Title: Effect of curcumin on glycerol-induced acute kidney injury in rats

    doi: 10.1038/s41598-017-10693-4

    Figure Lengend Snippet: Effect of curcumin on apoptotic, oxidative stress and lipid oxidative signaling pathway associated proteins in kidney tissue. ( a ) Representative western blots depicting protein levels of PI 3 K, p-PI 3 K Akt, p-Akt, AMPK, p-AMPK, cleaved caspase-3, cleaved caspase-9, Nrf2 and HO-1 in kidney tissue. ( b ) The CUR + AKI group showed significantly increased levels of p-PI 3 K, p-Akt, p-AMPK, Nrf2, and HO-1, and decreased levels of cleaved caspase-3, cleaved caspase-9 compared to the AKI and CO + AKI groups. Data are the means ± SD (n = 10). Statistical significance: *p

    Article Snippet: The membranes were blocked with 1× casein solution for approximately 4 h and then incubated with rabbit monoclonal anti-P-AMPK (Ser485) (2537), anti-P-Akt (Ser473) (4058), anti-Akt (4685), anti-cleaved caspase-3 (Asp175) (9664) and anti-Cleaved Caspase-9 (Asp330) (7237), all from Cell signaling technology; or rabbit polyclonal anti-Nrf2 (sc-722), anti-GAPDH (H-12) (sc-166574) and mouse monoclonal anti-HO-1 (sc-390991) anti-AMPK (sc-398861), all from Santa Cruz Biotechnology; or PI3K (ab86714) and p-PI3K (ab182651), both from Abcam Biotechnology, in blocking buffer for 2 h at room temperature.

    Techniques: Western Blot

    CAE-induced activation of NRF2/HO1 pathway was not linked with AHR activation. (a, b) HaCaT cells were transfected with control siRNA, AhR siRNA, or NrF2 siRNA using the DharmaFECT® Duo transfection reagent. After 24 h, the cells were incubated with CAE (100 μ g/ml) in the presence of B[a]P for 24 h and then subjected to real-time PCR analysis for HO-1 and CYP1A1 . ∗ p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Antagonizing Effects of Clematis apiifolia DC. Extract against Benzo[a]pyrene-Induced Damage to Human Keratinocytes

    doi: 10.1155/2019/2386163

    Figure Lengend Snippet: CAE-induced activation of NRF2/HO1 pathway was not linked with AHR activation. (a, b) HaCaT cells were transfected with control siRNA, AhR siRNA, or NrF2 siRNA using the DharmaFECT® Duo transfection reagent. After 24 h, the cells were incubated with CAE (100 μ g/ml) in the presence of B[a]P for 24 h and then subjected to real-time PCR analysis for HO-1 and CYP1A1 . ∗ p

    Article Snippet: The following are the cell culture reagents: B[a]P (CAS No. 50-32-8, purity 99.9%, Sigma-Aldrich Co., N.Y., USA), dasatinib (Src inhibitor, Sigma-Aldrich Co.), AhR antibodies (Santa Cruz Biotechnology, Santa Cruz, Ca, USA), ARNT antibodies (Santa Cruz Biotechnology), LaminB1 antibodies (Epitomic, Burlingame, CA, USA), α -tubulin antibodies (Epitomic), β -actin antibodies (Sigma-Aldrich Co.), phospho-Src (Tyr416) antibodies (Cell Signaling Technology Inc., Beverly, MA, USA), Src antibodies (Cell Signaling Technology Inc.), CYP1A1 antibodies (Santa Cruz Biotechnology), NQO1 antibodies (Santa Cruz Biotechnology), and HO-1 antibodies (Santa Cruz Biotechnology).

    Techniques: Activation Assay, Transfection, Incubation, Real-time Polymerase Chain Reaction