Journal: Experimental and Therapeutic Medicine

Article Title: Sinomenine activation of Nrf2 signaling prevents inflammation and cerebral injury in a mouse model of ischemic stroke

doi: 10.3892/etm.2021.10079

Figure Lengend Snippet: SINO treatment relieves MCAO-induced cerebral injuries. (A) Representative micrographs of H&E staining of brain sections (x20 magnification; scale bar, 50 µm). (B) Brain water content analysis in brain tissues from Sham (sham-operated, n=6), SINO (treatment only, n=6), MCAO (MCAO-operated, n=6) and SINO/MCAO (MCAO operated plus SINO treatment, n=6). (C) Western blot analysis of n-Nrf2, HO-1 and NQO1 protein expression levels from brain tissue. The samples from two randomly selected brains in each group were presented. (D) Quantification of brain protein expression levels. All the experiments were repeated at least three times. The data are presented as the mean ± SEM. * P<0.05 and ** P<0.01 vs. Sham; # P<0.05 and ## P<0.01 vs. MCAO. HO-1, heme oxygenase-1; MCAO, middle cerebral artery occlusion; n-Nrf2, nuclear-nuclear factor-erythroid 2-related factor; NQO1, NAD(P)H: Quinoneoxidoreductase 1; SINO, sinomenine.

Article Snippet: The following antibodies were used in the present study: SINO (MedChemExpress); ML385 (MedChemExpress); Nrf2, Keap1, NAD(P)H: Quinoneoxidoreductase (NQO1), Lamin B and HO-1 antibodies (Abcam); β-actin antibodies (Bioworld Technology, Inc.); NF-κB p65, Phosphorylated (p-)IκBα and IκBα antibodies (Cell Signaling Technology, Inc.); HRP-conjugated secondary antibodies (Bioworld Technology, Inc.); and Dylight488 conjugated goat anti-rabbit IgG secondary antibodies (Bioworld Technology, Inc.).

Techniques: Staining, Western Blot, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Sinomenine activation of Nrf2 signaling prevents inflammation and cerebral injury in a mouse model of ischemic stroke

doi: 10.3892/etm.2021.10079

Figure Lengend Snippet: SINO treatment activates the Nrf2 signaling pathway. (A) BV2 cells were treated with a serial of doses of SINO for 24 h. HO-1 and NQO1 protein expression levels were assayed using western blotting. (B) BV2 cells were treated with SINO (200 µM) for various durations. HO-1 and NQO1 protein expression levels were assayed using western blotting. (C) BV2 cells were treated with SINO (200 µM) for 4 h. n-Nrf2 and t-Nrf2 protein expression levels were assayed using western blotting. Nuclear protein Lamin B and β-actin were used as controls. (D) BV2 cells were treated with serial doses of SINO for 24 h. The mRNA expression levels of HO-1 and NQO1 were measured using reverse transcription-semi-quantitative PCR. (E) Cell viability assay. BV2 cells were treated with various doses of SINO for 24 h. The cell viability was assayed by a CCK-8 kit. All the experiments were repeated at least three times. * P<0.05 and ** P<0.01 vs. Control HO-1, heme oxygenase-1; NQO1, NAD(P)H: Quinoneoxidoreductase 1; n-, nuclear; Nrf2; nuclear factor-erythroid 2-related factor; SINO, sinomenine; t-, total.

Article Snippet: The following antibodies were used in the present study: SINO (MedChemExpress); ML385 (MedChemExpress); Nrf2, Keap1, NAD(P)H: Quinoneoxidoreductase (NQO1), Lamin B and HO-1 antibodies (Abcam); β-actin antibodies (Bioworld Technology, Inc.); NF-κB p65, Phosphorylated (p-)IκBα and IκBα antibodies (Cell Signaling Technology, Inc.); HRP-conjugated secondary antibodies (Bioworld Technology, Inc.); and Dylight488 conjugated goat anti-rabbit IgG secondary antibodies (Bioworld Technology, Inc.).

Techniques: Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Viability Assay, CCK-8 Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Sinomenine activation of Nrf2 signaling prevents inflammation and cerebral injury in a mouse model of ischemic stroke

doi: 10.3892/etm.2021.10079

Figure Lengend Snippet: SINO treatment regulates microglia polarization and inflammation in an Nrf2-dependent manner. BV2 cells were stimulated with OGD for 4 h followed by treatment with SINO (200 µM) for 12 h. The mRNA expression levels of (A) M1 markers (IL-6/NOS2) and (B) M2 markers (IL-10/Arg-1) were determined using RT-sqPCR. BV2 cells were pretreated with ML385 (5 µM) for 48 h to inhibit Nrf2 expression. Cells were then stimulated with OGD for 4 h followed by treatment with SINO (200 µM) for 12 h. The mRNA expression levels of (C) M1 and (D) M2 markers were measured using RT-sqPCR. The data are presented as the mean ± SEM of three independent experiments. * P<0.05 and ** P<0.01 vs. Control; ## P<0.01 vs. OGD. Arg-1, arginase-1; NOS2, nitric oxide synthase 2; OGD, oxygen and glucose deprivation; SINO, sinomenine; RT-sqPCR, reverse transcription-semi-quantitative PCR.

Article Snippet: The following antibodies were used in the present study: SINO (MedChemExpress); ML385 (MedChemExpress); Nrf2, Keap1, NAD(P)H: Quinoneoxidoreductase (NQO1), Lamin B and HO-1 antibodies (Abcam); β-actin antibodies (Bioworld Technology, Inc.); NF-κB p65, Phosphorylated (p-)IκBα and IκBα antibodies (Cell Signaling Technology, Inc.); HRP-conjugated secondary antibodies (Bioworld Technology, Inc.); and Dylight488 conjugated goat anti-rabbit IgG secondary antibodies (Bioworld Technology, Inc.).

Techniques: Expressing, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Experimental and Therapeutic Medicine

Article Title: Sinomenine activation of Nrf2 signaling prevents inflammation and cerebral injury in a mouse model of ischemic stroke

doi: 10.3892/etm.2021.10079

Figure Lengend Snippet: SINO treatment regulates microglia inflammation in an Nrf2-dependent manner. (A) BV2 cells were pretreated with ML385 (5 µM) for 48 h to inhibit Nrf2 expression. Cells were then stimulated with OGD for 4 h followed by treatment with SINO (200 µM) for 12 h. The protein expression levels of p-IκBα and total IκBα were analyzed using western blotting. (B) Immuno-fluorescent staining of cells was performed using an NF-κB p65 primary antibody and Dylight488 conjugated secondary antibody (middle row of panels). The cells were also stained with DAPI (top row of panels) and merged with NF-κB images (lower row of panels). The data are presented as the mean ± SEM of three independent experiments. ** P<0.01 vs. Control; ## P<0.01 vs. OGD. OGD, oxygen and glucose deprivation; p-, phosphorylated; SINO, sinomenine.

Article Snippet: The following antibodies were used in the present study: SINO (MedChemExpress); ML385 (MedChemExpress); Nrf2, Keap1, NAD(P)H: Quinoneoxidoreductase (NQO1), Lamin B and HO-1 antibodies (Abcam); β-actin antibodies (Bioworld Technology, Inc.); NF-κB p65, Phosphorylated (p-)IκBα and IκBα antibodies (Cell Signaling Technology, Inc.); HRP-conjugated secondary antibodies (Bioworld Technology, Inc.); and Dylight488 conjugated goat anti-rabbit IgG secondary antibodies (Bioworld Technology, Inc.).

Techniques: Expressing, Western Blot, Staining, Control

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: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: A) Kaplan-Meier survival analysis of mice hydrodynamically injected with the MYC transposon system and the indicated NRF2 activating guide RNAs (sgRNA) and Cas9; B) Representative diseased livers from mice injected with MYC and sgKeap1 or sgNrf2 (targeting the Keap1 interacting ETGE domain); C) In vivo growth of murine MYC/sgKeap1 tumors transduced with Nrf2-targeted or control shRNAs; Data from HCC lines 1 & 2 were combined and plotted (n=5 each); D) Kaplan-Meier survival analysis of mice that were hydrodynamically injected with the MYC transposon system and an sgRNA/Cas9 targeting Keap1 and subsequently treated with glutathione synthesis inhibitor BSO. (* indicates p-value < 0.05 by two-tailed student t test). See also Figure S1 and Table S1.

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: Injection, In Vivo, Transduction, Control, Two Tailed Test

Journal: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: A) Measuring NRF2 stability using an NRF2-nanoluciferase fusion protein in lysates of KEAP1 proficient HepG2 and KEAP1 mutant Huh1 cells transduced with control or shRNA against FN3K; mean of n=6 for HepG2 and n=3 for Huh1 cells ± SD; B) Relative expression of ~80 antioxidative genes in KEAP1N414Y mutant Huh1 cells transduced with control or FN3K shRNAs; average of four replicates (n=2 for each FN3K-specific shRNAs) relative to control shRNA; C) Unsupervised clustering of total proteomics data from indicated Huh1 cell lysates; GSEA analysis of over- and underrepresented proteins in FN3K-deficient cells shows reduction of NRF2 target proteins (top) and proteins involved in xenobiotic metabolism (bottom); D) Luminescence-based quantification of oxidized and reduced glutathione in KEAP1N414Y Huh1 cells expressing control vector or shRNA against FN3K; error bar represents SD from n ≥ 5 replicates; E) Nuclear extracts from Huh1 cells with control vector or FN3K knockdown immunoprecipitated with NRF2 or IgG antibodies and probed for MAFG and β-actin; nuclear lysates (bottom) were loaded on a separate gel and probed with αNRF2 and αLamin B as indicated; F) Schematic of KEAP1-dependent and independent mechanisms of NRF2 inhibition by glycation. (*denotes two-tailed t test calculated p-value of <0.05). See also Figure S4 and Table S4.

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: Mutagenesis, Transduction, Control, shRNA, Expressing, Plasmid Preparation, Knockdown, Immunoprecipitation, Inhibition, Two Tailed Test

Journal: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: A) Mass spectrometric identification of tryptic peptides from unglycated (upper) and in vitro glycated recombinant NRF2 (lower); peptides that decreased in abundance upon in vitro glycation are shown in black while those that are unaffected in red; B) AUC analysis of indicated tryptic peptides generated from non-glycated and in vitro glycated NRF2; data are represented relative to R25-R34 peptide and error bar represents SD from 3 replicates; C) Spectral plot of K487-R499 peptide from unmodified (left) and in vitro glycated NRF2 (right, 5 g/L glucose for 14 days); D) Spectral intensity graphs of indicated peptides obtained from tryptic digest of immunoprecipitated nuclear NRF2 from control and FN3K deficient Huh1 cells; E) Normalized AUC analysis of indicated tryptic peptides generated by digesting immunoprecipitated NRF2 from parental (n=2) or FN3K-silenced Huh1 cells (n=3) as indicated; error bar represents SD. (* indicates p-value < 0.05). See also Figures S5, S6, and Table S5.

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: In Vitro, Recombinant, Generated, Immunoprecipitation, Control

Journal: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: A) Diagram of dual gene-targeting strategy in murine HCCs; B) Ultrasound of murine livers 4 weeks after injection with indicated plasmids; tumors are marked in red; C) Ex vivo images of livers from animals injected with indicated sgRNA combinations; D) Subcutaneous Huh1 xenografts with and without FN3K knockdown and NAC treatment as indicated measured ~30 days after implantation; E) Decreased NRF2 target protein expression by immunoblot on the FN3K deficient Huh1 tumors from panel 5D; F) Phenyl borate enrichment and immunoblot shows NRF2 glycation in the FN3K deficient Huh1 xenografts. See also Figure S7 and Table S6.

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: Injection, Ex Vivo, Knockdown, Expressing, Western Blot

Journal: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: A) Diagram of our strategy for a genome-wide screen against NRF2 driven expression of the HSV-TK suicide gene; B) Map of the lentiviral vector directing ARE-controlled HSV-TK and luciferase expression; C) Change in sgRNA library representation comparing untreated cells and cells treated with the NRF2 inducer tBHQ and ganciclovir; D) Predicted sites of NRF2 protein glycation using indicated algorithms; TAD: Transactivation domain; E) Phenyl borate affinity purification and immunoblotting reveals NRF2 glycation upon FN3K knockdown in KEAP1 mutant Huh1 cells; values on top refers to % of glycated NRF2 represented by the ratio of NRF2 signal intensity in PB-bound (PB) to the sum PB-bound and flow through (FT); F) Immunoblot for nuclear (upper panel) and cytoplasmic (lower panel) levels of the indicated proteins in KEAP1 wild type HepG2 cells transduced and treated as indicated; G) Chromatin immunoprecipitation (ChIP) on indicated HepG2 nuclear lysates with anti-NRF2 antibody followed by amplification of indicated promoters; shown as % of input DNA and error bar is SD of 4 replicates; H) Viability of HepG2 cells untreated or treated with H2O2 (400 μM, 24 hours) with and with without pre-incubation with NAC (10 mM, 3 hours); mean of 9 replicates ± SD. (* indicates p-value < 0.05 by two-tailed student t test). See also Figure S3 and Table S3.

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: Genome Wide, Expressing, Plasmid Preparation, Luciferase, Affinity Purification, Western Blot, Knockdown, Mutagenesis, Chromatin Immunoprecipitation, Amplification, Incubation, Two Tailed Test

Journal: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: A) Unbiased, pan-cancer analysis of mutations that are significantly related (co-occurring or mutual exclusive) with KEAP1 and NRF2 using the SELECT algorithm (see text and methods for details); B) Oncoprint showing mutual exclusive relation between NRF2/ KEAP1 and EGFR mutations in human cancers; C) Nuclear extracts from HepG2 cells treated with NRF2 inducer DLS (1 μM, 24 hours), EGF (10 ng/mL, 24 hours), TGFα (1 nM, 24 hours) or DMSO and probed with antibodies against NRF2 and lamin B; D) Nuclear (upper panel) and cytoplasmic extracts (lower panel) from H3255 (EGFRL858R) cells transduced with EGFR-specific shRNA or control and immunoblotted with indicated antibodies; E) Viability of isogenic H3255 cells transduced with sgRNAs targeting KEAP1 or LacZ (control) and treated with erlotinib; error bars represent SD from 3 replicates; F) Lysates from paired PC9 cells transduced with indicated sgRNA-Cas9 constructs and treated with DMSO or erlotinib (10 nM, 6 hours) probed with the indicated antibodies. (* indicates p-value < 0.05 by two-tailed student t test). See also Figure S2 and Table S2.

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: Transduction, shRNA, Control, Construct, Two Tailed Test

Journal: Cell

Article Title: The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase

doi: 10.1016/j.cell.2019.07.031

Figure Lengend Snippet: GSE133160

Article Snippet: Recombinant human NRF2 , Origene , Cat#TP760529.

Techniques: Western Blot, Virus, Recombinant, Affinity Chromatography, SYBR Green Assay, Software, RNA Sequencing

Journal: Cell metabolism

Article Title: The Mitochondrial-Encoded Peptide MOTS-c Translocates to the Nucleus to Regulate Nuclear Gene Expression in Response to Metabolic Stress

doi: 10.1016/j.cmet.2018.06.008

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: NFE2L2 Human siRNA Oligo Duplex , Origene , Cat#SR321100.

Techniques: Recombinant, SYBR Green Assay, Reporter Assay, Over Expression, Sequencing, Luciferase, Software, Real-time Polymerase Chain Reaction

Journal: Cell metabolism

Article Title: The Mitochondrial-Encoded Peptide MOTS-c Translocates to the Nucleus to Regulate Nuclear Gene Expression in Response to Metabolic Stress

doi: 10.1016/j.cmet.2018.06.008

Figure Lengend Snippet:

Article Snippet: NFE2L2 Human siRNA Oligo Duplex , Origene , Cat#SR321100.

Techniques:

Journal: Metabolic Brain Disease

Article Title: The SIRT-1/Nrf-2/HO-1 antioxidant defense axis in adult attention-deficit/hyperactivity disorder

doi: 10.1007/s11011-026-01845-5

Figure Lengend Snippet: ROC Curve Analysis for HO-1, Nrf-2 and SIRT-1. HO-1 (cut-off value 0.87, sensitivity 78.3%, specificity 76.7%); Nrf-2 (cut-off value 9.8 sensitivity 70%, specificity 73.3%); SIRT-1 (cut-off value 8.3, sensitivity 75%, specificity 70%). Abbreviations: HO-1: heme oxygenases-1; Nrf-2: nuclear factor erythroid 2-related factor 2; SIRT-1: Sirtuin 1

Article Snippet: Serum HO-1, NRF-2, and SIRT-1 concentrations were analyzed using ELISA kits according to the manufacturers’ standard protocols (BT Lab, Human Heme Oxygenase-1: Cat. No. E0932Hu; BT Lab, Human Nuclear Factor Erythroid 2-Related Factor 2: Cat. No. E3244Hu; BT Lab, Human Sirtuin-1: Cat. No. E2557Hu; Jiaxing Korain Biotech, Jiaxing, China) on a Rel Assay automated ELISA reader (Biobase Biodusty Co., Ltd., Jinan, China).

Techniques:

Journal: Biomedical reports

Article Title: Mistimed H 2 S upregulation, Nrf2 activation and antioxidant proteins levels in renal tubular epithelial cells subjected to anoxia and reoxygenation.

doi: 10.3892/br.2020.1309

Figure Lengend Snippet: Figure 4. Expression of Nrf2 and its transcriptional targets under anoxia or reoxygenation. (A) Expression of Nrf2 and its transcriptional targets (B) SOD‑3, (C) glutathione reductase, (D) ferritin H and (E) xCT increased under anoxia, and returned to baseline levels during reoxygenation. Representative blots are presented. *P<0.001 vs. Ctrl; #P<0.001 vs. Reox. Nrf2, nuclear factor erythroid 2‑like 2; Ctrl, control; Reox, reoxygenation; OD, optical density; SOD‑3, super‑ oxide dismutase 3; xCT, cystine‑glutamate antiporter.

Article Snippet: Primary antibodies used were specific for CBS (1:1,000; cat. no. TA338394; OriGene Technologies Inc.), CSE (1:100; cat. no. sc‐374249; Santa Cruz Biotechnology, Inc.), 3‐MST (1:100; cat. no. sc‐376168; Santa Cruz Biotechnology, Inc.), Nrf2 (1:1,000; cat. no. TA343586; OriGene Technologies, Inc.), BIOMEDICAL REPORTS 13: 3, 2020 3 superoxide dismutase 3 (SOD3; 1:100; cat. no. sc‐271170; Santa Cruz Biotechnology, Inc.), glutathione reductase (GR; 1:100; cat. no. sc‐133245; Santa Cruz Biotechnology, Inc.), ferritin heavy chain (1:100; cat. no. sc‐376594; Santa Cruz Biotechnology, Inc.), cystine‐glutamate antiporter (xCT; 1:1,000; cat. no. ANT‐111; Alomone Labs), activated cleaved‐caspase‐3 (175) (1:500; cat. no. 7074; Cell Signaling Technology, Inc.), p53 (1:500; cat. no. 2524 Cell Signaling Technology, Inc.), p53 phosphorylated at serine 15 (p‐p53) (1:500; cat. no. 9284; Cell Signaling Technology, Inc.), Bax (1:500; cat. no. 5023; Cell Signaling Technology, Inc.) and β‐actin (1:2,500; cat. no. 4967; Cell Signaling Technology, Inc.).

Techniques: Expressing, Control