Journal: Open Medicine

Article Title: miR-141-3p accelerates ovarian cancer progression and promotes M2-like macrophage polarization by targeting the Keap1-Nrf2 pathway

doi: 10.1515/med-2023-0729

Figure Lengend Snippet: miR-141-3p promotes OC proliferation and M2 polarization via Keap1/Nrf2 pathway. (a) The miR-141-3p mimic promotes the proliferation of SKOV3 cells, while the promotional effect of miR-141-3p was abolished by Nrf2 inhibitor ML3508; (b) miR-141-3p mimic increases expression of M2-like macrophage biomarker CD206 and this effect is abolished by Nrf2 inhibitor; (c–f) The increasing mRNA levels of Arg1 and IL-10, and decreasing mRNA levels of iNOS and TNF-α caused by miR-141-3p mimic are reversed by Nrf2 inhibitor ML3508; (g) The protein levels of Keap1 and Nrf2 in SKOV3 cells after being treated by miR-141-3p NC, miR-141-3p mimic, and miR-141-3p mimic + ML3508.

Article Snippet: Nrf2 activator-2 (compound O15, HY-145879) was purchased from Med chem express (MCE, CA, USA), and this compound (5.0 μM) was added into the cell culture medium of SKOV3 followed by miR-141-3p inhibitor treatment.

Techniques: Expressing, Biomarker Assay

Journal: The Journal of Reproduction and Development

Article Title: Nrf2 inhibition affects cell cycle progression during early mouse embryo development

doi: 10.1262/jrd.2017-042

Figure Lengend Snippet: Effects of brusatol treatment on the expression of Nrf2. A) Images of control and brusatol treatment group by immunofluorescent staining. B) Nrf2 fluorescence intensity analysis of the control and brusatol treatment group. The fluorescence intensity of Nrf2 was significantly decreased in brusatol treatment group as compared with the control group. Green, Nrf2; blue, DNA. Scale bar, 20 µm. C) Nrf2 and UCHL1 protein expression after brusatol treatment by western blotting. D) Quantitative analysis of the relative expression of Nrf2 protein. Western blot analysis showing the decline in the expression of Nrf2 and UCHL1 in brusatol-treated group. * Significantly different as compared with the control and Nrf2 activation groups (P < 0.05). E) Nrf2 mRNA expression in brusatol treatment group as compared with the control group.

Article Snippet: The cytoplasm of pronucleus-stage zygotes was microinjected with Nrf2 CRISPR activation plasmid (sc-421869-ACT; Santa Cruz Biotechnology, San Jose, CA, USA) to induce Nrf2 protein overexpression.

Techniques: Expressing, Control, Staining, Fluorescence, Western Blot, Activation Assay

Journal: The Journal of Reproduction and Development

Article Title: Nrf2 inhibition affects cell cycle progression during early mouse embryo development

doi: 10.1262/jrd.2017-042

Figure Lengend Snippet: Effects of brusatol treatment on Nrf2-mediated oxidative stress. Brusatol reduced the mRNA level of Nrf2 target genes, HO-1 , GCLC , and SOD1 , as analyzed by qRT-PCR.

Article Snippet: The cytoplasm of pronucleus-stage zygotes was microinjected with Nrf2 CRISPR activation plasmid (sc-421869-ACT; Santa Cruz Biotechnology, San Jose, CA, USA) to induce Nrf2 protein overexpression.

Techniques: Quantitative RT-PCR

Journal: The Journal of Reproduction and Development

Article Title: Nrf2 inhibition affects cell cycle progression during early mouse embryo development

doi: 10.1262/jrd.2017-042

Figure Lengend Snippet: Effects of microinjection of Nrf2 CRISPR activation plasmid in the cytoplasm of the zygote on the embryo development potency. A) Embryos were microinjected with Nrf2 CRISPR activation plasmid in a medium supplemented with brusatol for 24, 48, 72 and 96 h. Nrf2 activation group showed a significant increase in the four-cell stage embryos as compared with brusatol treatment group. B) Microinjection with Nrf2 CRISPR activation plasmid partially rescued the embryo development potency. The embryo development rate from four-cell to blastocyst stage was significantly increased as compared with the embryos treated with brusatol. As shown in , the expression of Nrf2 protein was increased in Nrf2 activation group as compared with the brusatol treatment group. a, b, and c represent the significant differences in three groups. Original magnification × 200.

Article Snippet: The cytoplasm of pronucleus-stage zygotes was microinjected with Nrf2 CRISPR activation plasmid (sc-421869-ACT; Santa Cruz Biotechnology, San Jose, CA, USA) to induce Nrf2 protein overexpression.

Techniques: Microinjection, CRISPR, Activation Assay, Plasmid Preparation, Expressing

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Protective effects of sulforaphane in experimental vascular cognitive impairment: Contribution of the Nrf2 pathway

doi: 10.1177/0271678X18764083

Figure Lengend Snippet: Sfn reduces neuronal death in vivo and in vitro. Six weeks after 2VO, brain sections at dorsal hippocampus were collected and stained with LFB-CV. (a) Representative images of the parietal cortex (upper panels, scale bar = 100 µm) and hippocampal CA1 (lower panels, scale bar = 200 µm). (b, c) Neurons with normal-like morphology were counted and data were expressed as number per mm2. 2VO reduced normal-like neurons in both cortex and CA1, which was partially reversed by Sfn. n = 6, *p < 0.05, **p < 0.01, ***p < 0.001 vs. Sham; ##p < 0.01, ###p < 0.001 vs. 2VO. Nrf2 knockdown in rat primary neurons was induced by sh-Nrf2 transfection, and OGD was performed with or without Sfn pretreatment. (d) Hoechst 33258 staining with cell counting and (e) LDH assay showed that Sfn ameliorated OGD-induced cell death, which was abolished by Nrf2 knockdown. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Ctrl; #p < 0.05, ##p < 0.01 vs. OGD; &p<0.05 vs. OGD+Sfn.

Article Snippet: In vitro Nrf2 knockdown To knock down Nrf2 in neurons and MBMECs, lentiviral particles carrying shRNA targeting Nrf2 (sh-Nrf2) or a scramble sequence (sh-Sc) were added to cells together with 8 μg/ml polybrene (all from Santa Cruz, Dallas, TX) for 16 h. The virus-containing culture media were then replaced with fresh growth media.

Techniques: In Vivo, In Vitro, Staining, Knockdown, Transfection, Cell Counting, Lactate Dehydrogenase Assay

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Protective effects of sulforaphane in experimental vascular cognitive impairment: Contribution of the Nrf2 pathway

doi: 10.1177/0271678X18764083

Figure Lengend Snippet: Sfn protects the BBB via Nrf2 activation and TJ preservation. MBMECs were subjected to OGD with or without Sfn pretreatment. (a) Live/dead staining (scale bar = 50 µm) with cell counting and (b) LDH assay showed that Sfn protected against OGD-induced cell death. (c) In an in vitro BBB model, Sfn partially reversed OGD-induced BBB leakage. ***p < 0.001 vs. Ctrl; #p < 0.05, ##p < 0.01, ###p < 0.001 vs. OGD. Nrf2 was knocked down by sh-Nrf2 transfection. (d) Live/dead staining (scale bar = 50 µm) with cell counting and (e) LDH assay revealed that Sfn-provided protection was abolished by Nrf2 knockdown. n.s.: not significant, **p < 0.01. (f) In vitro BBB permeability assessment without OGD showed that Nrf2 knockdown disrupted the BBB integrity. *p < 0.05, **p < 0.01, ***p < 0.001 vs. sh-Sc. (g) Dual-luciferase assay revealed that CLD5 promoter activity was decreased by Nrf2 knockdown. n.s.: not significant, **p < 0.01, ***p < 0.001. (h) Six weeks after 2VO, the brain sections were collected and immunohistochemistry showed that Sfn reduced CLD5 loss in the cortex (scale bar = 20 µm). (i–k) Representative Western blots and semi-quantitative analysis showed that Sfn reduced loss of CLD5 and occludin after 2VO. *p < 0.05 vs. Sham; #p < 0.05, ##p < 0.01 vs. 2VO.

Article Snippet: In vitro Nrf2 knockdown To knock down Nrf2 in neurons and MBMECs, lentiviral particles carrying shRNA targeting Nrf2 (sh-Nrf2) or a scramble sequence (sh-Sc) were added to cells together with 8 μg/ml polybrene (all from Santa Cruz, Dallas, TX) for 16 h. The virus-containing culture media were then replaced with fresh growth media.

Techniques: Activation Assay, Preserving, Staining, Cell Counting, Lactate Dehydrogenase Assay, In Vitro, Transfection, Knockdown, Permeability, Luciferase, Activity Assay, Immunohistochemistry, Western Blot

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Protective effects of sulforaphane in experimental vascular cognitive impairment: Contribution of the Nrf2 pathway

doi: 10.1177/0271678X18764083

Figure Lengend Snippet: Sfn activates Nrf2 and upregulates HO-1 expression after 2VO in rats. Cortical tissues were harvested at indicated time-points, and nuclear and cytosolic proteins were extracted. (a) Representative Western blots and semi-quantitative analysis within one week after 2VO, showing that 2VO-induced Nrf2 activation was enhanced by Sfn administration. (b) Representative Western blots and semi-quantitative at six weeks after 2VO, showing long-lasting activation of Nrf2 pathway by Sfn. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Sham; #p < 0.05, ##p < 0.01 vs. 2VO. (c) Representative images of double-labeling of HO-1 with cellular markers (NeuN for neurons, GFAP for astrocytes and microvessels, Iba-1 for microglia/macrophages, and APC for mature oligodendrocytes) revealed wide-spread HO-1 expression after 2VO. Scale bar = 50 µm.

Article Snippet: In vitro Nrf2 knockdown To knock down Nrf2 in neurons and MBMECs, lentiviral particles carrying shRNA targeting Nrf2 (sh-Nrf2) or a scramble sequence (sh-Sc) were added to cells together with 8 μg/ml polybrene (all from Santa Cruz, Dallas, TX) for 16 h. The virus-containing culture media were then replaced with fresh growth media.

Techniques: Expressing, Western Blot, Activation Assay, Labeling

Journal: The FASEB Journal

Article Title: Sabotaging of the oxidative stress response by an oncogenic noncoding RNA

doi: 10.1096/fj.201600654R

Figure Lengend Snippet: NRF2 response to ACA11 generated ROS and levels of antioxidants. A) Western blot of nuclear and cytosolic fractions showing earlier translocation of NRF2 to nucleus in ACA11 cells in response to H2O2 (left). Bar graphs (right) show densitometry of bands from left panel. B) Activation of NRF2 is abrogated in presence of ACA11. Results of NRF2 reporter assay at baseline and in presence of H2O2 stimulus. C, D) ChIP analysis of 2 NRF2 target genes, HMXO-1 (C) and TXN-1 (D), in MM1.S cells with vector (pLKO.1) and ACA11 overexpression. E–G) qPCR analysis of specific NRF2 target genes HMXO-1 (E), TXN-1 (F), and PRDXN1 (G) in presence or absence of hydrogen peroxide in MM1.S cells, respectively. Representative figures are shown of experiments completed at least 3 times in triplicate. All data points represent means ± sd (n = 3). Asterisks indicate significance according to 1-way ANOVA with Bonferroni posttest. *P < 0.05, **P < 0.01, ***P < 0.001 compared to pLKO.1 in MM1.S.

Article Snippet: Nuclear protein (10 μg) was used to determine the NRF2 binding activity using the TransAM NRF2 ELISA kit (Active Motif) according to the manufacturer’s guidelines.

Techniques: Generated, Western Blot, Translocation Assay, Activation Assay, Reporter Assay, Plasmid Preparation, Over Expression

Journal: PLoS ONE

Article Title: Nrf2-Inducing Anti-Oxidation Stress Response in the Rat Liver - New Beneficial Effect of Lansoprazole

doi: 10.1371/journal.pone.0097419

Figure Lengend Snippet: List of oligonucleotide primers used for RT-PCR.

Article Snippet: Mixtures of plant extracts, such as “Protandim” (LifeVantage, USA) and “Nrf2 activator” (XYMOGEN, USA) are available as inducers of the Keap1/Nrf2/ARE system.

Techniques: Derivative Assay

Journal: PLoS ONE

Article Title: Nrf2-Inducing Anti-Oxidation Stress Response in the Rat Liver - New Beneficial Effect of Lansoprazole

doi: 10.1371/journal.pone.0097419

Figure Lengend Snippet: A) Up-regulation of Nrf2 mRNA in the liver at 3 h. ** P<0.01, ***P<0.001 compared with control. B) Up-regulation of Nrf2 immunoreactivity (IR) in the liver at 6 h. *P<0.05, ** P<0.01 compared with control. Representative photographs showing an increase of Nrf2 IR. C) No significant changes in Keap1 mRNA levels in the liver at 3 h. D) No significant changes in Keap1 IR levels in the liver at 6h. Representative photographs showing no change of Keap1 IR. E) Nuclear translocation of Nrf2 in the liver demonstrated by western blotting of cytosol fraction (C; Calpain-positive) and nuclear fraction (N; Histone H1-positive) at 3h and 6h. F) Nuclear translocation of Nrf2 in hepatocytes as demonstrated by double fluorescence immunohistochemistry. Blue indicates DAPI-positive nuclei, red indicates Nrf2 IR, and pink indicates the nuclear localization of Nrf2. Bar = 20 µm.

Article Snippet: Mixtures of plant extracts, such as “Protandim” (LifeVantage, USA) and “Nrf2 activator” (XYMOGEN, USA) are available as inducers of the Keap1/Nrf2/ARE system.

Techniques: Control, Translocation Assay, Western Blot, Fluorescence, Immunohistochemistry

Journal: PLoS ONE

Article Title: Nrf2-Inducing Anti-Oxidation Stress Response in the Rat Liver - New Beneficial Effect of Lansoprazole

doi: 10.1371/journal.pone.0097419

Figure Lengend Snippet: A) No significant changes in the levels of mRNA for Nrf2, Keap1 and HO-1. B) Up-regulation of IR levels for Nrf2 and HO-1, with no change of Keap1 IR. *P<0.05, ***P<0.001 compared with control. Representative photographs showing an increase of Nrf2 and HO-1 IR. C) Nuclear translocation of Nrf2 in hepatocytes as demonstrated by double fluorescence immunohistochemistry and western blotting of cytosol fraction (C; Calpain-positive) and nuclear fraction (N; Histone H1-positive). Blue indicates DAPI-positive nuclei, red indicates Nrf2 IR, and pink indicates the nuclear localization of Nrf2. Bar = 20 µm.

Article Snippet: Mixtures of plant extracts, such as “Protandim” (LifeVantage, USA) and “Nrf2 activator” (XYMOGEN, USA) are available as inducers of the Keap1/Nrf2/ARE system.

Techniques: Control, Translocation Assay, Fluorescence, Immunohistochemistry, Western Blot

Journal: PLoS ONE

Article Title: Nrf2-Inducing Anti-Oxidation Stress Response in the Rat Liver - New Beneficial Effect of Lansoprazole

doi: 10.1371/journal.pone.0097419

Figure Lengend Snippet: A summary schematic of the lansoprazole-induced AhR/Cyp1a1/Nrf2 pathway in the liver. XRE, xenobiotic response elements; ARE, antioxidant response element.

Article Snippet: Mixtures of plant extracts, such as “Protandim” (LifeVantage, USA) and “Nrf2 activator” (XYMOGEN, USA) are available as inducers of the Keap1/Nrf2/ARE system.

Techniques:

Journal: Cancer Cell International

Article Title: Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes

doi: 10.1186/s12935-023-03056-9

Figure Lengend Snippet: Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type. Violin plots of TMB ( E ) and mutation count ( F ) among four groups: Nrf2-activating mutations, Nrf2-inactivating mutations, unknown NFE2L2 mutations and WT groups. TMB: tumor mutation burden. WT: wide type

Article Snippet: Fig. 2 Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type.

Techniques: Mutagenesis

Journal: Cancer Cell International

Article Title: Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes

doi: 10.1186/s12935-023-03056-9

Figure Lengend Snippet: Summary of NFE2L2 Nrf2-activating mutations in the MSK MetTropism cohort. ( A ) Distribution of NFE2L2 mutations according to locations of the mutation. ( B ) The prevalence of Nrf2-activating mutations and Nrf2-inactivating mutations in each cancer type. ( C ) Kaplan–Meier survival curves of OS among Nrf2-activating mutations, Nrf2-inactivating mutations, unknown NFE2L2 mutations and WT groups. ( D ) Association of Nrf2-inactivating mutations with OS stratified by cancer type. ( E ) The multivariate Cox regression analysis for OS in patients. MSI: microsatellite instability; FGA: fraction genome altered; OS: overall survival; MU: mutation; WT: wild type

Article Snippet: Fig. 2 Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type.

Techniques: Mutagenesis

Journal: Cancer Cell International

Article Title: Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes

doi: 10.1186/s12935-023-03056-9

Figure Lengend Snippet: Clinical outcomes for Nrf2-activating mutations in NSCLC patients from TCGA cohort. ( A ) Kaplan–Meier survival curves of OS between NFE2L2 MU and WT groups from LUAD. datasets. ( B ) The prevalence of Nrf2-activating mutations and Nrf2-inactivating mutations in each cancer type. ( C ) The proportion of Nrf2-activating mutations and Nrf2-inactivating mutations in LUAD and LUSC. Kaplan–Meier survival curves of OS among Nrf2-activating MU, Nrf2-inactivating MU, unknown NFE2L2 MU and WT groups from LUAD ( D ) and LUSC ( E ). ( F ) Kaplan–Meier survival curves of OS among HTMB NFE2L2 MU, HTMB NFE2L2 WT, LTMB NFE2L2 MU and LTMB NFE2L2 WT patients with NSCLC from TCGA cohort. HTMB: high tumor mutation burden, L TMB: Low tumor mutation burden. ( G ) The multivariate Cox regression analysis for OS in patients with LUAD. CNA: copy number alteration; MSI: microsatellite instability; OS: overall survival

Article Snippet: Fig. 2 Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type.

Techniques: Mutagenesis

Journal: Cancer Cell International

Article Title: Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes

doi: 10.1186/s12935-023-03056-9

Figure Lengend Snippet: Summary of NFE2L2 mutations and survival outcomes in patients receiving ICIs. ( A ) Kaplan–Meier survival curves of OS between NFE2L2 MU and NFE2L2 WT groups in the pooled cohort composed of DFCI, MSK, MSK1661, OAK, Pender, POPLAR and PUCH cohorts. ( B ) Association of NFE2L2 mutation with OS stratified by study with ICIs treatment. The proportion of CR/PR to ICIs in NFE2L2 mutated patients from the pooled ( C , n = 908) and NSCLC ( G , n = 491) cohorts. Kaplan–Meier survival curves of OS among Nrf2-activating MU, Nrf2-inactivating MU, unknown NFE2L2 MU and WT groups from pan cancer ( D ) and NSCLC ( H ) with ICIs treatment. ( E) The multivariate Cox regression analysis for OS in patients with ICIs treatment. ( F ) Meta-analysis for NSCLC and bladder cancer to summarize association of NFE2L2 mutation with OS after ICIs treatment

Article Snippet: Fig. 2 Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type.

Techniques: Mutagenesis

Journal: Cancer Cell International

Article Title: Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes

doi: 10.1186/s12935-023-03056-9

Figure Lengend Snippet: Significantly enriched signaling pathways in NFE2L2-MU ( A ), Nrf2-activating MU ( B ) and Nrf2-inactivating MU patients with NSCLC in TCGA cohort. GSEA: gene set enrichment analysis; NES: normalized enrichment scores

Article Snippet: Fig. 2 Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type.

Techniques: Protein-Protein interactions

Journal: Cancer Cell International

Article Title: Pan-cancer analysis of NFE2L2 mutations identifies a subset of lung cancers with distinct genomic and improved immunotherapy outcomes

doi: 10.1186/s12935-023-03056-9

Figure Lengend Snippet: Landscape of the microenvironment phenotypes in NSCLC. ( A ) Heatmap representation of TME subtype Fges from TCGA NSCLC cohort using unsupervised hierarchical clustering. Distribution of Nrf2-activating MU and Nrf2-inactivating MU according to four TME subtypes ( B ) and five immune subtypes ( C ) in TCGA NSCLC cohort. ( D ) Immune deconvolution using xCell enriched in Nrf2-activating MU and Nrf2-inactivating MU patients with NSCLC. ( E ) Violin plots of TCR Richness among Nrf2-activating mutations, Nrf2-inactivating mutations, unknown NFE2L2 mutations and WT groups

Article Snippet: Fig. 2 Overview of NFE2L2 mutations and Nrf2-activating mutations in the OrigiMed cohort. ( A ) The NFE2L2 mutations profiles and the clinical characters of 10,194 patients. ( B ) Distribution of NFE2L2 mutations according to locations of the mutation. ( C ) Co-occurrence/mutual exclusivity of NFE2L2 and other genes. ( D ) The prevalence of Nrf2-activating mutations, Nrf2-inactivating mutations and unknown NFE2L2 mutations in each cancer type.

Techniques:

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Effects of radiocontrast on nuclear factor, erythroid 2 like 2 (Nrf2) and cleaved caspase 3 expression. Representative western blot analyses of Nrf2 and cleaved caspase 3 expression over time after iohexol treatment and quantitative analyses are shown: in vitro ( A , B ) and in vivo ( C , D ) experiments. The expression of Nrf2 was significantly increased after iohexol treatment. Cropped gels are used in the figure, and the full-size gels are presented in Supplementary Figures – . * p < 0.05 vs . Cont and # p < 0.05 vs . CM 1 h. Abbreviations: Cont, saline control group; CM, iohexol treatment group.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: Expressing, Western Blot, In Vitro, In Vivo, Saline, Control

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Changes in creatinine levels, histologic findings in the kidneys and renal injury markers after iohexol treatment. ( A ) Increased serum creatinine levels after iohexol treatment were aggravated by the loss of Nrf2 function. *p < 0.05 vs . WTcon and KOcon, # p < 0.05 vs . WT_CM. ( B ) Representative histology of the tubular changes after contrast injury. Iohexol treatment in mice induced tubular dilatation and vacuolization with tubular structure disruption. The severity and extent of tubular injury were aggravated upon the loss of Nrf2 (periodic acid-Schiff (PAS) staining; original magnification, ×400). ( C ) Quantitative measurement of tubular injury in histology by measuring the area of tubular dilatation and vacuolization. The area of tubular injury after iohexol treatment was significantly increased in the group with Nrf2 loss compared to that of the other groups. *p < 0.05 vs. WTcon and KOcon, # p < 0.05 vs . WT_CM. ( D ) Immunohistochemical staining of NGAL. Iohexol treatment in mice increased NGAL staining area and intensity in tubules, and these effects were further aggravated by the loss of Nrf2 (NGAL; Original magnification, ×400). ( E ) The area of NGAL positive staining was calculated and compared between groups. The histology of iohexol-treated Nrf2 KO mice showed significantly increased NGAL staining in renal tubules compared to that of the other groups. *p < 0.05 vs. WTcon and KOcon, # p < 0.05 vs . WT_CM. Abbreviations: WTcon, wild-type control group; KOcon, Nrf2 knockout control group; WT_CM, wild-type iohexol-treated group; KO_CM, Nrf2 knockout iohexol-treated group; Nrf2, nuclear factor, erythroid 2 like 2.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: Disruption, Staining, Immunohistochemical staining, Control, Knock-Out

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Cell cytotoxicity assays after iohexol treatment and Nrf2 inhibition. Nrf2 inhibition aggravated the decrease in cell viability measured by MTT assay after iohexol treatment. * p < 0.05 vs . Cont and siRNA, # p < 0 . 05 vs . CM. Abbreviations: Cont, control group; siRNA, siRNA transfection group; CM, iohexol-only treatment group, CM + siRNA, iohexol-treated group with Nrf2 inhibition by siRNA transfection; Nrf2, nuclear factor, erythroid 2 like 2; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: Inhibition, MTT Assay, Control, Transfection

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Loss of Nrf2 in mice enhanced iohexol-induced apoptosis. Terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling (TUNEL) staining of kidney tissue ( A ) and quantification of TUNEL-positive apoptotic nuclei (B) revealed an increase in apoptosis in renal tubules after iohexol treatment. In addition, this effect was aggravated by the loss of Nrf2. * p < 0.05 vs. WTcon and KOcon, # p < 0.05 vs. WT_CM. ( C ) Representative immunoblot analysis and quantitative analysis showed increased cleaved caspase 3 expression in tubular cells after iohexol treatment. Inhibition of Nrf2 enhanced cleaved caspase 3 expression. Cropped gels are used in the figure, and the full-size gels are presented in Supplementary Figure . * p < 0.05 vs . Cont and siRNA, # p < 0 . 05 vs . CM. Abbreviations: WTcon, wild-type control group; KOcon, Nrf2 knockout control group; WT_CM, wild-type iohexol-treated group; KO_CM, Nrf2 knockout iohexol-treated group; Cont, control group; siRNA, siRNA transfection group; CM, iohexol-only treatment group, CM + siRNA, iohexol treated group with Nrf2 inhibition by siRNA transfection; Nrf2, nuclear factor, erythroid 2 like 2.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: End Labeling, TUNEL Assay, Staining, Western Blot, Expressing, Inhibition, Control, Knock-Out, Transfection

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Loss of Nrf2 enhanced the increase in iohexol-induced oxidative stress. ( A ) Oxidative stress in the kidneys determined by MDA concentration was increased after iohexol treatment, and this effect was augmented in mice with the loss of Nrf2 function. * p < 0.05 vs . WTcon and KOcon, # p < 0.05 vs . WT_CM. ( B ) ROS production according to DCF-DA staining was observed by confocal microscopy (2000× magnification). Green-colored immunofluorescence was increased after iohexol treatment, and this effect was enhanced by Nrf2 inhibition. ( C ) The percentage of fluorescence intensity of DCF-stained cells measured by a fluorescence spectrophotometer showed similar results. * p < 0.05 vs . Cont and siRNA, # p < 0 . 05 vs . CM 3 h and 6 h. Abbreviations: WTcon, wild-type control group; KOcon, Nrf2 knockout control group; WT_CM, wild-type iohexol-treated group; KO_CM, Nrf2 knockout iohexol-treated group; Cont, control group; siRNA, siRNA transfection group; CM, iohexol-only treatment group, CM + siRNA, iohexol-treated group with Nrf2 inhibition by siRNA transfection; Nrf2, nuclear factor, erythroid 2 like 2; DCF-DA, dichlorofluorescein diacetate.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: Concentration Assay, Staining, Confocal Microscopy, Immunofluorescence, Inhibition, Fluorescence, Spectrophotometry, Control, Knock-Out, Transfection

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Immunoblot analyses of HO-1 and cytochrome c . An increase in HO-1 expression after iohexol treatment was not observed upon reduced Nrf2 function in both in vivo and in vitro experiments ( A ). Reduced Nrf2 function was also associated with enhanced cytochrome c expression. Cropped gels are used in the figure, and the full-size gels are presented in Supplementary Figure . ( B ). *p < 0.05 vs . Cont/siRNA or WTcon/KOcon. # p < 0.05 vs. CM or WT_CM. Cropped gels are used in the figure, and the full-size gels are presented in Supplementary Figure . Abbreviations: Cont, control group; siRNA, siRNA transfection group; CM, iohexol-only treated group, CM + siRNA, iohexol-treated group with Nrf2 inhibition by siRNA transfection; WTcon, wild-type control group; KOcon, Nrf2 knockout control group; WT_CM, wild-type iohexol-treated group; KO_CM, Nrf2 knockout iohexol-treated group; Nrf2, nuclear factor, erythroid 2 like 2.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: Western Blot, Expressing, In Vivo, In Vitro, Control, Transfection, Inhibition, Knock-Out

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Loss of Nrf2 promotes macrophage infiltration in the renal tubules after iohexol treatment. Immunohistochemical staining of F4/80 ( A ) and quantitative analysis of F4/80 positive cells (red arrow, ( B ) showed increased macrophage infiltration after iohexol treatment, and these effects were augmented by the loss of Nrf2 function. * p < 0.05 vs . WTcon and KOcon, # p < 0.05 vs . WT_CM. Abbreviations: WTcon, wild-type control group; KOcon, Nrf2 knockout control group; WT_CM, wild-type iohexol-treated group; KO_CM, Nrf2 knockout iohexol-treated group; Nrf2, nuclear factor, erythroid 2 like 2.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: Immunohistochemical staining, Staining, Control, Knock-Out

Journal: Scientific Reports

Article Title: The role of nuclear factor erythroid-2-related factor 2 expression in radiocontrast-induced nephropathy

doi: 10.1038/s41598-019-39534-2

Figure Lengend Snippet: Quantitative analysis of cell viability by 5-(3-carboxymethoxyphenyl)-2H-tetrazolium inner salt (MTS) assay after iohexol and Nrf2 activator (CDDO-Me) co-treatment with or without the inhibition of Nrf2 expression by siRNA transfection in NRK-52E cells. Cell viability was not changed upon pretreatment of CDDO itself. However, decreased cell viability after iohexol treatment was significantly attenuated by Nrf2 activation with 6-hour CDDO pretreatment. Nrf2 inhibition by siRNA pretreatment enhanced the decrease in cell viability after iohexol treatment, and CDDO pretreatment partially recovered the cell viability decrease induced by iohexol and Nrf2 inhibition * p < 0.05 vs . Con, CDDO, tcon, and tcon + CDDO, # p < 0.05 vs . CM, and † p < 0.05 vs . CM + CDDO, ‡ p < 0.05 vs . tCM.

Article Snippet: NRK-52E cells were treated with the Nrf2 activator bardoxolone methyl (CDDO-Me; Toronto Research Chemicals, Toronto, ON, Canada) to confirm the role of Nrf2 in iohexol-induced tubular injury.

Techniques: MTS Assay, Inhibition, Expressing, Transfection, Activation Assay