Journal: Journal of Veterinary Science

Article Title: Epigallocatechin-3-gallate suppresses hemin-aggravated colon carcinogenesis through Nrf2-inhibited mitochondrial reactive oxygen species accumulation

doi: 10.4142/jvs.22097

Figure Lengend Snippet: WST, water-soluble tetrazolium salt; EGCG, epigallocatechin-3-gallate; Nrf2, nuclear factor erythroid-2-related factor 2. * p < 0.05.

Article Snippet: Nrf2 (NBP1-32822) and Keap1 (NBP2-03319) antibodies were purchased from Novus Biologicals (Centennial, CO, USA).

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: SIRT1 and NRF2 Gene Transfer Mediate Distinct Neuroprotective Effects Upon Retinal Ganglion Cell Survival and Function in Experimental Optic Neuritis

doi: 10.1167/iovs.17-22972

Figure Lengend Snippet: Design and in vitro characterization of AAV2 vectors. (A) Outline of AAV expression cassettes used in the study. RT-qPCR analysis of relative quantities of human NRF2 or human SIRT1 mRNA in 84-31 cells treated with (B) AAV2-NRF2 and (C) AAV2-SIRT1 compared to nontransduced cells. Micrographs displaying vector protein expression following ARPE-19 cell transduction with (D) AAV2-eGFP, (E) AAV2-NRF2, and (F) AAV2-SIRT1. (G) Control micrograph of ARPE-19 cells transduced with AAV2-NRF2 without anti-FLAG primary antibody incubation.

Article Snippet: Human SIRT1 (transcript variant 1) and human NRF2 (transcript variant 1) cDNA clones were obtained from Origene.

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Plasmid Preparation, Transduction, Control, Incubation

Journal: Investigative Ophthalmology & Visual Science

Article Title: SIRT1 and NRF2 Gene Transfer Mediate Distinct Neuroprotective Effects Upon Retinal Ganglion Cell Survival and Function in Experimental Optic Neuritis

doi: 10.1167/iovs.17-22972

Figure Lengend Snippet: AAV2 transduction profile and RGC transduction efficiency following intravitreal delivery. (A) Representative micrograph of retinal flatmount following intravitreal injection of AAV2-eGFP. RGCs are labeled with Brn3a (red). (B) Representative visual field of a retinal flatmount used for calculating RGC transduction efficiency with AAV2. (C) Quantification of RGC transduction (n = 5 retina). (D) Representative cross-section of mouse retina following intravitreal injection of AAV2-eGFP, (E) AAV2-NRF2, and (F) AAV2-SIRT1. RGCs are labeled with Brn3a (red). Cells positively transduced with NRF2 or SIRT1 vectors are labeled with FLAG (green). Data represented as mean ± SEM.

Article Snippet: Human SIRT1 (transcript variant 1) and human NRF2 (transcript variant 1) cDNA clones were obtained from Origene.

Techniques: Transduction, Injection, Labeling

Journal: Investigative Ophthalmology & Visual Science

Article Title: SIRT1 and NRF2 Gene Transfer Mediate Distinct Neuroprotective Effects Upon Retinal Ganglion Cell Survival and Function in Experimental Optic Neuritis

doi: 10.1167/iovs.17-22972

Figure Lengend Snippet: Effect of gene transfer on visual acuity during EAE. OKR recordings demonstrate significantly decreased visual acuity in eyes of EAE mice treated with vehicle (n = 1 0) or AAV2-eGFP (n = 10). Treatment with AAV2-NRF2 (n = 25) did not improve visual function. Mice treated with AAV2-SIRT1 (n = 25) show trending improvement in OKR at days 28 to 49 compared to EAE eyes injected with vehicle and significant improvement at days 35 (P = 0.032) and 42 (P = 0.049) compared to AAV2-eGFP injected eyes also subjected to EAE. Data represented as mean ± SEM. *P < 0.05, **P < 0.01 by 1-way ANOVA with Tukey's HSD post-test.

Article Snippet: Human SIRT1 (transcript variant 1) and human NRF2 (transcript variant 1) cDNA clones were obtained from Origene.

Techniques: Injection

Journal: Food Science & Nutrition

Article Title: Trans‐Fatty Acids ( TFA ) Induced Vascular Injury Through the Regulation of the Sirt1‐Ppargc1a‐Nfe2l2 Signaling Pathway in Male Rats

doi: 10.1002/fsn3.70975

Figure Lengend Snippet: Effect of trans‐fatty acids on Sirt1/Ppargc1a/Nfe2l2 pathway of rat blood vessels. (A) Western blotting results of Silent information regulator 1 (Sirt1), Peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (Ppargc1a), and nuclear factor erythroid 2‐related factor 2 (Nfe2l2). (B−D) Semi‐quantitative analyses of protein expression. Results are shown as mean ± SD ( n = 6). * p < 0.05 by contrast with ND group; # p < 0.05 by contrast with LTD group. ND, normal diet; LTD, 1% trans‐fatty acids diet; HTD, 4% trans‐fatty acids diet.

Article Snippet: The following primary antibodies were employed in the study: anti‐rabbit Bcl2 (Signalway Antibody, China), anti‐rabbit Bax (Cell Signaling Technology, USA), anti‐rabbit Ppargc1a (Affinity, China), anti‐rabbit Casp3 (GeneTex, USA), anti‐rabbit Cleaved Casp3 (Arigo Biolaboratories, China), anti‐rabbit Nfe2l2 (Medical & Biological Laboratories, China), anti‐rabbit Sirt1 (OriGene, USA), and anti‐rabbit β‐actin (Abcam, China).

Techniques: Western Blot, Expressing

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 1. High expression of Nrf2 associates with AML relapse. A, analysis of gene expression profile of Nrf2 in AML samples (n = 173) and matched normal samples (n = 70) according to GEPIA website. B, the expression level of Nrf2 mRNA in normal (n = 17), complete remission (n = 33), and relapsed AML patients (n = 46) was identified by RT–PCR. C, the expression of Nrf2 was detected by RT–PCR in the same AML patients before and after relapse (n = 8). D, Western blot was adopted for detecting the expression level of Nrf2 in normal (n = 9), complete remission (n = 9), and relapsed (n = 9) AML samples.

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Expressing, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 2. High expression of OGG1 associates with high expression of Nrf2 in AML. A–D, RT–PCR analysis of the expressions of OGG1, XRCC1, AP site endonuclease 1 (APE1), and DNA polymerase β (POL-β) in Nrf2-low (n = 29) and Nrf2-high (n = 26) groups. E, the gene expression profiles of OGG1 in AML samples (n = 173) and matched normal samples (n = 70) were analyzed according to GEPIA website. F, RT–PCR analysis of OGG1 expression in normal (n = 28), complete remission (n = 29), and relapsed (n = 26) AML patients. G, the expression of OGG1 in the same AML patients before and after relapse was

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 3. High expression of Nrf2 in Ara-C-resistant AML cell lines. A, U937 and U937R cells were treated with different concentrations of Ara-C (4, 16, 64, 192, 386, 578, 768, and 1536 μM) for 24 h, and the cell viability was detected by CCK-8 colorimetry. B, THP-1 and THP-1R cells were treated with different concentrations of Ara-C (0.5, 4, 64, 192, 386, 578, 768, and 1536 μM) for 24 h, and the cell viability was detected by CCK-8 colorimetry. C, after U937, U937R, THP-1, and THP-1R were cocultured with Ara-C (2 μM) for 24 h, the apoptosis rate of AML cell lines was detected by FCM. D, analysis of apoptosis rate.

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Expressing, CCK-8 Assay, Colorimetric Assay

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 4. High expression of Nrf2 decreases the sensitivity of leukemic cells to Ara-C. A, Western blot was used to verify the upregulation of Nrf2 in AML- sensitive celllines (U937andTHP-1). B, the relative grayvalues of Nrf2expression. C, Western blotwas appliedtodetect thedownregulation ofNrf2in AML-resistant cell lines (U937R andTHP-1R). D, the relativegray values of Nrf2 expression. E, RT–PCR verifiedthe expression ofmRNA after Nrf2 regulation.F, FCM was employed todetect the apoptosis rate of AML-sensitive cell lines cocultured with Ara-C (2 μM) for 24 h after Nrf2 overexpression. G, after downregulation of Nrf2, the apoptosis rate of AML drug–resistant cell lines cocultured with Ara-C (100 μM) for 24 h was detected by FCM. Each experiment was repeated three times independently and expressed as mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001. AML, acute myeloid leukemia; Ara-C, cytarabine; FCM, flow cytometry; Nrf2, nuclear factor E2–related factor 2.

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Over Expression, Cytometry

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 5. Overexpression of Nrf2 promotes the expression of OGG1 to mediate drug resistance in AML cells. A, detection of OGG1 expression in normal (bone marrow mononuclear cells from normal healthy donors) and AML-sensitive cell lines (U937 and THP-1) and drug-resistant cell lines (U937R and THP-1R) by IF staining. Scale bars represent 20 μm. B, quantification of OGG1 relative fluorescence intensity in normal and leukemia cells by ImageJ. C, Western blot was employed to detect OGG1 protein expression after upregulation of Nrf2. D, the relative gray values of Nrf2 and OGG1 expressions.

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Over Expression, Expressing, Staining, Western Blot

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 7. Nrf2 overexpression promotes OGG1 expression by activating AKT signaling pathway. A, GeneMania protein–protein interaction network of OGG1, Nrf2, and AKT. B and C, Western blot was applied to identify the expressions of Nrf2, OGG1, AKT, p-AKT, Cleaved-caspase 9, and Total-caspase 9 proteins in AML-sensitive cell lines after overexpression of Nrf2. D and E, Western blot was adopted for detecting the expression of Nrf2, OGG1, AKT, p-AKT, Cleaved-caspase 9, and Total-caspase 9 proteins in AML drug–resistant cell lines after downregulation of Nrf2. Each experiment was repeated three times independently and denoted as mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001. Nrf2, nuclear factor E2–related factor 2; OGG1, 8-hydroxyguanine DNA glycosidase.

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Over Expression, Expressing, Western Blot

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 8. Inhibition of AKT signaling pathway to reduce OGG1 expression. A, after U937 cells were treated with 2 μM MK-2206 for 24 h, the protein expression levels of Nrf2, OGG1, AKT, p-AKT, Cleaved-caspase 9, and Total-caspase 9 in Nrf2 overexpression and EV1 groups were detected by Western blot. B, the relative gray value of related proteins. C, after THP-1 cells were treated with 2 μM MK-2206 for 24 h, the protein expression levels of Nrf2, OGG1, AKT, p-AKT, Cleaved-caspase 9, and Total-caspase 9 in Nrf2 overexpression and EV1 groups were detected by Western blot. D, the relative gray values of related

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: Inhibition, Expressing, Over Expression, Western Blot

Journal: The Journal of biological chemistry

Article Title: Nuclear factor Nrf2 promotes glycosidase OGG1 expression by activating the AKT pathway to enhance leukemia cell resistance to cytarabine.

doi: 10.1016/j.jbc.2022.102798

Figure Lengend Snippet: Figure 9. Downregulation of Nrf2 reduces the risk of drug resistance in vivo. A and B, tumor volume of mice after subcutaneous injection of U937R (n = 4), U937R-EV (n = 4), U937R-si-Nrf2 (n = 3), and U937R-EV + Ara-C (n = 4) and U937R-si-Nrf2 + Ara-C (n = 3) cells after 35 days growth. C, changes of tumor weight after subcutaneousinjectionofU937R(n=4),U937R-EV(n=4),U937R-si-Nrf2(n=3),andU937R-EV+Ara-C(n=4)andU937R-si-Nrf2+Ara-C(n=3)cells.DandE,volumegrowth curveofxenografttumor.F,analysisanddrawingofsurvivalcurveofxenograftedtumormicebyKaplan–Meiermethod.G–J,IHCwasappliedtoidentifytheexpressionsof Nrf2andOGG1insubcutaneoustransplantedtumortissuesections.Thescalebarsfromlefttorightrepresent100and50μm,respectively.*p< 0.05,**p< 0.01,and***p< 0.001. Ara-C, cytarabine; EV, empty vector; IHC, immunohistochemistry; Nrf2, nuclear factor E2–related factor 2; OGG1, 8-hydroxyguanine DNA glycosidase.

Article Snippet: The Nrf2 antibody was acquired from the abcam, and the OGG1 antibody was purchased from the Novus.

Techniques: In Vivo, Injection, Plasmid Preparation, Immunohistochemistry

Journal: Aging

Article Title: Pharmacological recapitulation of the lean phenotype induced by the lifespan-extending sulfur amino acid-restricted diet.

doi: 10.18632/aging.206237

Figure Lengend Snippet: Figure 3. The SAAR diet and BSO exert tissue-specific effects on Nrf2 and Phgdh. The SAAR diet increased Nrf2 (A) and Phgdh (B) protein expressions in the liver, which ultimately resulted in higher serine concentrations (C). Unlike the SAAR diet, BSO did not increase Nrf2 and Phgdh in the liver but increased both in the kidneys (D, E). Regardless of the changes in Nrf2 and Phgdh, BSO increased serine concentrations in livers and kidneys (C–F). NAC reversed SAAR-induced changes in Nrf2, Phgdh, and serine (A–F). Note: Sample size = 5-6. Statistical methods are similar to those in Figure 2.

Article Snippet: Host species Antibody dilutions Primary Antibodies Nrf2 Cell Signaling Technology 20733 Rabbit 1:1000 in 5% BSAa Phgdh Cell Signaling Technology 13428 Rabbit 1:1000 in 5% milka β-Actin Sigma A5441 Mouse 1:15000 in 5% milka Vinculin Proteintech 66305-1-Ig Mouse 1:10000 in 5% milka Secondary Antibodies Anti-Rabbit-HRP Cell Signaling Technology 7074 Goat 1:4000 in 5% milk, 1hb Anti-Mouse-HRP Bio-Rad 170-6516 Goat 1:20000 in 5% milk, 30 minb aAll incubations were performed overnight at 4° C. bAll incubations were performed at room temperature. www.aging-us.com 22 AGING Supplementary Table 3.

Techniques:

Journal: Redox Biology

Article Title: Considerations for antibody-based detection of NRF2 in human cells

doi: 10.1016/j.redox.2025.103549

Figure Lengend Snippet: NRF2 antibodies targeting different epitopes detect 3 bands migrating between 100 and 130 kDa in 8% Tris-glycine SDS-PAGE. (A) NRF2 migration in Tris-glycine 8 % SDS-PAGE in H1299, RERF and A549 cells at steady state and upon treatment with tert-butylhydroquinone (t-BHQ). (B) Knockdown of NRF2 gene ( NFE2L2 ) with a pool of siRNA targeting NFE2L2 (siNRF2) compared to transfection with control unspecific short RNAs (ctrl) in H1299 and RERF cells. (C,D) NRF2 dephosphorylation with λ phosphatase in RERF and H1299 cell lysates. Lysates were incubated with or without λ phosphatase for 30 min in 30 °C in the presence of MnCl 2 and NRF2 was detected by western blot. Arrows indicate bands detected by anti-NRF2 antibodies from Abcam [EP1808Y] (C) and Cell Signaling [D1Z9C] (D). (E) Detection of NRF2 in cytoplasmic and nuclear fractions upon treatment with tert-BHQ in H1299, RERF and A549 cells. Arrows indicate signal detected by anti-NRF2 antibodies from Abcam [EP1808Y]. Alfa-tubulin is a marker of cytoplasmic fraction and lamin is a marker of nuclear fraction. (F) Pulse-chase experiment upon translation inhibition with emetine in RERF cells. Cells were treated with emetine for indicated time and NRF2 levels were detected with Abcam [EP1808Y] antibodies. Molecular Weight is depicted at each western blot.

Article Snippet: NRF2 was detected with anti-NRF2 antibodies from Cell Signaling (E5F1A).

Techniques: SDS Page, Migration, Knockdown, Transfection, Control, De-Phosphorylation Assay, Incubation, Western Blot, Marker, Pulse Chase, Inhibition, Molecular Weight

Journal: Redox Biology

Article Title: Considerations for antibody-based detection of NRF2 in human cells

doi: 10.1016/j.redox.2025.103549

Figure Lengend Snippet: Calmegin is precipitated by anti-NRF2 antibodies and co-migrates with NRF2 in SDS-PAGE. (A) A scheme representing the workflow of identification of proteins precipitated by anti-NRF2 antibodies in RERF cells were treated with translation inhibitor emetine for 2 h cells or with tert-BHQ for 5 h. NRF2 was precipitated with Abcam EP1808Y antibodies. Precipitates were resolved in 8 % Tris-glycine SDS-PAGE and stained with Flamingo Fluorescent Gel Stain. Gel pieces with proteins migrating in 100–130 kDa range, representing NRF2 signal in western blot, were excised, followed by in-gel tryptic digestion of proteins. Diagrams show peptides of NRF2 (A) and calmegin (B) detected with LC-MS/MS in each sample (in duplicates). No NRF2 peptides were detected upon translation inhibition with emetine.

Article Snippet: NRF2 was detected with anti-NRF2 antibodies from Cell Signaling (E5F1A).

Techniques: SDS Page, Staining, Western Blot, Liquid Chromatography with Mass Spectroscopy, Inhibition

Journal: Redox Biology

Article Title: Considerations for antibody-based detection of NRF2 in human cells

doi: 10.1016/j.redox.2025.103549

Figure Lengend Snippet: Various commercial monoclonal anti-NRF2 antibodies bind calmegin. Upon calmegin (CLGN) knockdown in RERF (A) and H1299 (B) cells, NRF2 was detected with three different monoclonal anti-NRF2 antibodies: Abcam EP1808Y, Cell Signaling D1Z9C and ABclonal ARC0806. All these three antibodies bind to calmegin. (C) Anti-NRF2 antibody from Cell Signaling (E5F1A) binds little calmegin in RERF or no detectable calmegin in H1299 cells. (D) RERF cells were knocked down for NRF2 expression (siNRF2) or transfected with control short RNA (Ctrl) for 48 h and treated with NRF2 activator tert-BHQ (tBHQ). NRF2 was detected with anti-NRF2 antibodies from Cell Signaling (E5F1A).

Article Snippet: NRF2 was detected with anti-NRF2 antibodies from Cell Signaling (E5F1A).

Techniques: Knockdown, Expressing, Transfection, Control

Journal: Redox Biology

Article Title: Considerations for antibody-based detection of NRF2 in human cells

doi: 10.1016/j.redox.2025.103549

Figure Lengend Snippet: NRF2 domain structure with marked localization of immunogens used for the production of monoclonal antibodies used in this study, marked by a wave (∼), based on the information provided by a manufacturer. Antibodies which bind NRF2 with the highest specificity (Cell Signaling E5F1A) are marked with blue. Cell Signaling E5F1A - immunogen is a synthetic peptide corresponding to residues surrounding Ile86 ; Cell Signaling D1Z9C - immunogen is a synthetic peptide corresponding to residues surrounding Ala275 ; Abcam EP1808Y - immunogen is a synthetic peptide surrounding Leu550 ; ABclonal A3577 - immunogen is a synthetic peptide corresponding to a sequence within amino acids 505–605.

Article Snippet: NRF2 was detected with anti-NRF2 antibodies from Cell Signaling (E5F1A).

Techniques: Bioprocessing, Sequencing

Journal: Redox Biology

Article Title: Considerations for antibody-based detection of NRF2 in human cells

doi: 10.1016/j.redox.2025.103549

Figure Lengend Snippet: Analysis of selected monoclonal anti-NRF2 antibodies specificity in immunofluorescence in H1299 cells. Cells were knocked down for NRF2 expression (siNRF2) or transfected with control short RNA (Ctrl) for 48 h and, where indicated, treated with tert-BHQ (tBHQ) for 5 h. After that cells were fixed with 4 % para-formaldehyde and permabilized with 0.2 % Triton X-100, followed by staining with various anti-NRF2 antibodies (A,D,E) or analyzed for NRF2 levels in western blot (B). Alternatively, non-transfected cells were stained with anti-calmegin (CLGN) antibodies (C). Nuclei were stained with DAPI. All experiments were performed in at least three independent biological repeats with two technical repeats for each condition.

Article Snippet: NRF2 was detected with anti-NRF2 antibodies from Cell Signaling (E5F1A).

Techniques: Immunofluorescence, Expressing, Transfection, Control, Staining, Western Blot

Journal: Scientific Reports

Article Title: RNA sequencing of corneas from two keratoconus patient groups identifies potential biomarkers and decreased NRF2-antioxidant responses

doi: 10.1038/s41598-020-66735-x

Figure Lengend Snippet: Canonical pathways significantly altered in KC and KJ. Grey bars show total number of genes in the pathway (scale on top X axis), with blue and red bars showing the numbers of down-regulated and up-regulated genes (scale on bottom X axis). The p value next to each bar was calculated for that pathway by the IPA Core Analysis. “Acute Phase” is the Acute Phase Response Signaling pathway; “NRF2-mediated” is NRF2-mediated Oxidative Stress Response pathway; “ECM related” is the Hepatic Fibrosis / Hepatic Stellate Cell Activation pathway; “Rheumatoid Arthritis” is the Role of Macrophages, Fibroblasts and Endothelial Cells in Rheumatoid Arthritis pathway; and, “Osteoarthritis” includes genes associated with osteoarthritis.

Article Snippet: The following primary antibodies were used from Biorbyt Research Products: NRF2 (orb128433, 1:50), KEAP1 (orb48426, 1:100), Hsp40 (orb520080, 1:100), and GAS1 (orb414757, 1:100) and RXRA antibody were used from Novus Biologicals (NBP2–75653, 1:50).

Techniques: Activation Assay

Journal: Scientific Reports

Article Title: RNA sequencing of corneas from two keratoconus patient groups identifies potential biomarkers and decreased NRF2-antioxidant responses

doi: 10.1038/s41598-020-66735-x

Figure Lengend Snippet: NRF2 target genes significantly altered in KCN corneas. ( A ) Decreased NRF2 target gene expression in KC and KJ RNA seq. ( B ) Mechanism of NRF2 regulation by KEAP1; the latter binds to NRF2 and CUL3 for NRF2-ubiquitination and degradation to maintain low levels of NRF2 under homeostatic conditions. Under oxidative stress, KEAP1 dissociates from NRF2 allowing its increase and upregulation of target genes.

Article Snippet: The following primary antibodies were used from Biorbyt Research Products: NRF2 (orb128433, 1:50), KEAP1 (orb48426, 1:100), Hsp40 (orb520080, 1:100), and GAS1 (orb414757, 1:100) and RXRA antibody were used from Novus Biologicals (NBP2–75653, 1:50).

Techniques: Targeted Gene Expression, RNA Sequencing, Ubiquitin Proteomics

Journal: Scientific Reports

Article Title: RNA sequencing of corneas from two keratoconus patient groups identifies potential biomarkers and decreased NRF2-antioxidant responses

doi: 10.1038/s41598-020-66735-x

Figure Lengend Snippet: KEAP1 and NRF2 immunostaining in DN and KCN corneas. ( A ) KEAP1 staining is decreased in KCN corneas, with focally increased staining in some basal epithelial cells (inset), whereas in DN corneas KEAP1 shows staining of all epithelial layers (inset). ( B ) NRF2 shows very little to no staining of KCN corneas and these were all cytoplasmic (inset), while DN sections show stronger staining of epithelial cells and some nuclear staining (inset) DAPI nuclear staining shown in blue. IF staining of additional KCN and DN cornea sections are shown in Supplemental Fig. . Scale bar: 50 µm.

Article Snippet: The following primary antibodies were used from Biorbyt Research Products: NRF2 (orb128433, 1:50), KEAP1 (orb48426, 1:100), Hsp40 (orb520080, 1:100), and GAS1 (orb414757, 1:100) and RXRA antibody were used from Novus Biologicals (NBP2–75653, 1:50).

Techniques: Immunostaining, Staining