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: 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: Free radical biology & medicine

Article Title: NAD(P)H:quinone oxidoreductase 1 activity reduces hypertrophy in 3T3-L1 adipocytes.

doi: 10.1016/j.freeradbiomed.2012.05.047

Figure Lengend Snippet: Fig. 2. Keap1 protein decreases in differentiated adipocytes, while steady-state mRNA expression for Nrf2 and Keap1 increases. 3T3-L1 cells were differentiated as described under Materials and methods and harvested on Days 0, 4, 8, 11, and 14. (A) Nrf2 and (C) Keap1 (70 kDa) protein expression was examined by immunoblotting. Black bars, full-length (FL) Nrf2 (56 kDa); gray bars, degraded (Deg) Nrf2 (40 kDa); white bars, total (Tot) Nrf2; the sums of full-length Nrf2 and degraded Nrf2 were determined to obtain relative total Nrf2 protein expression. (B) Relative steady-state Nrf2 and (D) Keap1 mRNA levels were measured by SYBRGreen qPCR and normalized to 18S rRNA. Significance was determined by one-way ANOVA Dunnett’s for protein and one-way ANOVA Bonferroni’s for mRNA; * Pr0.05 as compared to Day 0, bars indicate Pr0.05 between samples. Data are expressed as mean7SD; n¼3–4.

Article Snippet: We also tried the Santa Cruz N-terminal Nrf2 antibody (No. sc-13032) that gave 6–7 signals ranging from 30 to 100 kDa and another C-terminal Nrf2 antibody from R & D Systems (No. MAB3925) that gave 20þ signals ranging from 20 to 100þ kDa (data not shown).

Techniques: Expressing, Western Blot

Journal: Free radical biology & medicine

Article Title: NAD(P)H:quinone oxidoreductase 1 activity reduces hypertrophy in 3T3-L1 adipocytes.

doi: 10.1016/j.freeradbiomed.2012.05.047

Figure Lengend Snippet: Fig. 3. NQO1 protein increases during limited clonal expansion and postmitotic growth arrest of differentiation. 3T3-L1 cells were differentiated as described under Materials and methods and harvested on Days 0, 1, 2, 3, and 4. (A) NQO1 and (D) Keap1 protein expressions were examined by immunoblotting. (B) Relative NQO1, (C) Nrf2, and (E) Keap1 mRNA levels were measured by SYBRGreen qPCR and normalized to 18S rRNA. Significance was determined by one-way ANOVA Dunnett’s for protein and one-way ANOVA Bonferroni’s for mRNA; * Pr0.05 as compared to Day 0, bars indicate Pr0.05 between samples. Data are expressed as mean7SD; n¼3–4.

Article Snippet: We also tried the Santa Cruz N-terminal Nrf2 antibody (No. sc-13032) that gave 6–7 signals ranging from 30 to 100 kDa and another C-terminal Nrf2 antibody from R & D Systems (No. MAB3925) that gave 20þ signals ranging from 20 to 100þ kDa (data not shown).

Techniques: Western Blot

Journal: Free radical biology & medicine

Article Title: NAD(P)H:quinone oxidoreductase 1 activity reduces hypertrophy in 3T3-L1 adipocytes.

doi: 10.1016/j.freeradbiomed.2012.05.047

Figure Lengend Snippet: Fig. 4. NQO1 and Keap1 proteins, but not mRNA, decreases as adipocyte differentiation progresses. 3T3-L1 cells were differentiated as described under Materials and methods and harvested on Days 4, 5, 6, 7, and 8. (A) NQO1 and (D) Keap1 protein expressions were examined by immunoblotting. (B) Relative NQO1, (C) Nrf2, and (E) Keap1 mRNA levels were measured by SYBRGreen qPCR and normalized to 18S rRNA. Significance was determined by one-way ANOVA Dunnett’s for protein and one-way ANOVA Bonferroni’s for mRNA; * Pr0.05 as compared to Day 4. Data are expressed as mean7SD; n¼3.

Article Snippet: We also tried the Santa Cruz N-terminal Nrf2 antibody (No. sc-13032) that gave 6–7 signals ranging from 30 to 100 kDa and another C-terminal Nrf2 antibody from R & D Systems (No. MAB3925) that gave 20þ signals ranging from 20 to 100þ kDa (data not shown).

Techniques: Western Blot

Journal: Free radical biology & medicine

Article Title: NAD(P)H:quinone oxidoreductase 1 activity reduces hypertrophy in 3T3-L1 adipocytes.

doi: 10.1016/j.freeradbiomed.2012.05.047

Figure Lengend Snippet: Fig. 6. LiCl treatment increases NQO1 protein, but not NQO1 mRNA. 3T3-L1 cells were differentiated as described under Materials and methods, and treated with varying concentrations of LiCl at Day 8 and harvested on Day 11. A 40 mM NaCl was used as a vehicle control (VC). (A) NQO1 was examined by immunoblotting. (B) Relative NQO1 and (C) Nrf2 mRNA levels were measured by SYBRGreen qPCR and normalized to 18S rRNA. Significance was determined by one-way ANOVA Dunnett’s; * Pr0.05 as compared to VC for protein and mRNA. Data are expressed as mean7SD; n¼3.

Article Snippet: We also tried the Santa Cruz N-terminal Nrf2 antibody (No. sc-13032) that gave 6–7 signals ranging from 30 to 100 kDa and another C-terminal Nrf2 antibody from R & D Systems (No. MAB3925) that gave 20þ signals ranging from 20 to 100þ kDa (data not shown).

Techniques: Control, Western Blot

Journal: Free radical biology & medicine

Article Title: NAD(P)H:quinone oxidoreductase 1 activity reduces hypertrophy in 3T3-L1 adipocytes.

doi: 10.1016/j.freeradbiomed.2012.05.047

Figure Lengend Snippet: Fig. 7. Sulforaphane increases NQO1 protein and mRNA levels. 3T3-L1 cells were differentiated as described under Materials and methods, and treated with varying concentrations of sulforaphane (SFN). A 0.3% DMSO was used as a VC. (A) NQO1 was examined by immunoblotting. (B) Relative NQO1 and (C) Nrf2 mRNA levels were measured by SYBRGreen qPCR and normalized to 18S rRNA. Significance was determined by one-way ANOVA Dunnett’s; * Pr0.05 as compared to VC for protein and mRNA. Data are expressed as mean7SD; n¼3.

Article Snippet: We also tried the Santa Cruz N-terminal Nrf2 antibody (No. sc-13032) that gave 6–7 signals ranging from 30 to 100 kDa and another C-terminal Nrf2 antibody from R & D Systems (No. MAB3925) that gave 20þ signals ranging from 20 to 100þ kDa (data not shown).

Techniques: 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 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