Journal: Journal of Oral Biology and Craniofacial Research

Article Title: Salivary defense system alters in vegetarian

doi: 10.1016/j.jobcr.2013.05.004

Figure Lengend Snippet: Superoxide dismutase activity in salivary fluid of vegetarian compared to non-vegetarian group (p < 0.05 significant).

Article Snippet: Superoxide dismutase (SOD) assay The activity was measured using a special superoxide dismutase kit made by R&D Systems Europe, Ltd. for research purposes, Catalog number: 7500-100-K.

Techniques: Activity Assay

Journal: Cancer gene therapy

Article Title: Targeting SOD1 via RNAi with PEGylated graphene oxide nanoparticles in platinum-resistant ovarian cancer.

doi: 10.1038/s41417-023-00659-2

Figure Lengend Snippet: Fig. 3 Nanoparticle behaviour in biological solutions. A Colloidal stability of GO, GOPEI, and GOPEI-mPEG were assessed in DIW, 0.9% NaCl, and RPMI + 10% FBS, 1 h after treatment. Nanoparticle aggregation was observed in all three biological solutions containing non- functionalized GO. B, C Gel-electrophoresis-based serum stability was carried out in GOPEI1x, GOPEI2x, and GOPEI3x. The susceptibility of siRNA to RNase degradation in serum was assessed for 48 h. Naked siSCR was used as a control. D Kinetic study of nanocarrier-siRNA serum stability was measured using mouse serum in siSCRFAM conjugated GOPEI, GOPEI-mPEG, and Lipo2000 relative to control naked siSCRFAM. The fluorescence of siSCRFAM was measured over a time of up to 8 h. E Kinetic study of nanoparticle behaviour was conducted in vitro using siSCRFAM complexed GO, GOPEI and GOPEI-mPEG. The degree of quenching induced by GOPEI upon binding siSCRFAM was measured by a Varioskan fluorescent plate reader. A 10 h time-course study of the live cell uptake of the nanoparticles was measured by recording their fluorescent emission. Samples containing only cells and RPMI media were used as negative controls. F Confocal laser scanning images showing the cellular uptake of GOPEI- siSCRFAM. Merged fluorescence images of cells treated with GOPEI- siSCRFAM with a magnified image of a single A2780 cell showing successful transfection of GOPEI as aggregations were compared with Lipo2000 as the positive control, and naked siSCRFAM was used as a negative control. G Quantitative assessment of nanoparticle: siRNA cellular uptake GOPEI-siRNA transfection efficiency was analyzed using flow cytometry. Two siSCRFAM concentrations of 60 nM and 90 nM complexed with Lipo2000 served as positive control. H Image showing intracellular graphene accumulation in cell pellets following three washing steps and centrifugation prior to protein isolation. I, J Time-course study showing SOD1 mRNA and protein levels following SOD1 knockdown. K, L The effect of concentration dependent SOD1 knockdown on protein levels was determined by Western blot and mRNA expression levels were measured using RT-qPCR. All values are expressed as mean ± SD. ns- not significant, *p < 0.05, **p < 0.01, and ****p < 0.0001 as analyzed using two-tailed unpaired t-test.

Article Snippet: The membrane was incubated with mouse anti-human SOD1 (15.9 kDa) monoclonal antibody (Cat# 67480-1-Ig, 1:1000, ProteinTech, Rosemont, IL, USA) and with mouse β-actin (42 kDa) monoclonal antibody (1:2000, Cat# 66009-1-Ig, ProteinTech) overnight at 40 °C.

Techniques: Nucleic Acid Electrophoresis, Control, In Vitro, Binding Assay, Transfection, Positive Control, Negative Control, Cytometry, Centrifugation, Isolation, Knockdown, Concentration Assay, Western Blot, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Cancer gene therapy

Article Title: Targeting SOD1 via RNAi with PEGylated graphene oxide nanoparticles in platinum-resistant ovarian cancer.

doi: 10.1038/s41417-023-00659-2

Figure Lengend Snippet: Fig. 4 In vitro nanoparticle toxicity. A Schematic diagram of the in vitro induction of acquired platinum resistance. B Baseline cisplatin sensitivity of A2780 and A2780DDP cell lines. C The IC50 of A2780 and A2780DDP were 4.71 ± 0.26 and 13.95 ± 1.18 µg/ml, respectively. D Bar graph showing normalized baseline SOD1 mRNA levels (n = 6). E Western blot showing the SOD1 protein levels in A2780 and A2780DDP cell lines in vivo in Q1 and Q3. F Bar graph showing normalized baseline SOD1 mRNA (n = 3). G Schematic illustration of xenograft tumour sample preparation for immunoblotting and qRT-PCR. H Western blot showing the SOD1 protein levels in A2780 and A2780DDP cell-derived xenograft tumour tissue samples. I Cisplatin treatment of A2780DDP cells (n = 6) induced SOD1 mRNA overexpression in a concentration-dependent manner measured by RT-qPCR. J Time-course evaluation SOD1 mRNA induction by GOPEI treatment in A2780DDP cells (n = 6) measured by qRT-PCR. K Cytotoxicity of GO, PEI, GOPEI and GOPEI-mPEG were determined at the following concentrations: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35 and 40 µg/mL, respectively. L MTT assay showing relative cell viability after A2780 cells were treated with 9 µg/mL of GOPEI for 48 h followed by cisplatin treatment at 14 different concentrations (2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 28 µg/mL respectively). IC50 values were compared with that of A2780 cells (n = 6). M Volcano plot showing the transcriptional activation of the mitochondrial unfolded protein response by 15 µM cisplatin in A2780 cell line. N, O GOPEI and GOPEI-mPEG treatment-induced differential in vitro activation of the UPRmt in (N) A2780 and (O) A2780DDP cell lines. P Schematic diagram illustrating UPRmt activation by cisplatin, graphene and cationic polymers leading to mitochondrial dysfunction and subsequent mito-nuclear signalling.

Article Snippet: The membrane was incubated with mouse anti-human SOD1 (15.9 kDa) monoclonal antibody (Cat# 67480-1-Ig, 1:1000, ProteinTech, Rosemont, IL, USA) and with mouse β-actin (42 kDa) monoclonal antibody (1:2000, Cat# 66009-1-Ig, ProteinTech) overnight at 40 °C.

Techniques: In Vitro, Western Blot, In Vivo, Sample Prep, Quantitative RT-PCR, Derivative Assay, Over Expression, Concentration Assay, MTT Assay, Activation Assay

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 3. VAC alleviated the renal inflammatory response and oxidative stress in T2DM mice (A) MDA contents in diabetic kidney tissues. n=6. (B) GSH-Px activity in diabetic kidney tissues. n=6. (C) Averaged fluorescence intensity of DHE fluorescence in diabetic kidney tissues. n=6. (D) Averaged fluorescence intensity of DCFH-DA fluorescence staining of diabetic kidney tissues. n=6. (E) DHE fluorescence staining or DCFH-DA fluorescence staining of diabetic kidney tissues. Scale bar: 100 μm. (F) F4/80 staining of diabetic kidney tissues. Scale bar: 50 μm, and the average fluorescence intensity of F4/80-expressing diabetic kidney tissues is shown. (G‒M) Representative blot images and quantitative analysis of phosphorylated NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. n=4. *P<0.05, **P<0.01, ***P<0.001 vs Ctrl. #P<0.05, ##P<0.01, ###P<0.001 vs DN.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Activity Assay, Fluorescence, Staining, Expressing

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 5. VAC alleviated the inflammatory response and oxidative stress in diabetic kidneys HK-2 cells were preincubated with 5 μM VAC for 12 h, followed by exposure to 35 mM HG for 48 h. (A–C) Relative mRNA levels of IL-1β, VCAM-1 and COX2. (D-F) DHE fluorescence staining or DCFH-DA fluorescence staining of HK-2 cells. Scale bar: 100 μm. (G–K) Relative mRNA levels of Nrf2, catalase, SOD3, SOD2 and SOD1. (L–R) Representative blot images and quantitative analysis of phosphorylated NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. *P<0.05, **P<0.01, ***P<0.001 vs NG. #P<0.05, ##P<0.01, ###P<0.001 vs HG. n=4.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Fluorescence, Staining

Journal: Acta biochimica et biophysica Sinica

Article Title: Vaccarin suppresses diabetic nephropathy through inhibiting the EGFR/ERK1/2 signaling pathway.

doi: 10.3724/abbs.2024141

Figure Lengend Snippet: Figure 8. VAC ameliorated fibrosis, the inflammatory response and oxidative stress in HG-induced HK-2 cells exposed to the EGFR inhibitor AG1478 or the ERK inhibitor U0126 (A–D) Relative mRNA levels of collagen-1, TGF-β1, α-SMA and E-cadherin in HK-2 cells. (E–G) Relative mRNA levels of IL-1β, VCAM-1 and COX-2 in HK-2 cells. (H) Averaged fluorescence intensity of DHE fluorescence in HK-2 cells. (I) DHE staining was performed on HG-induced HK-2 cells. Scale bar: 100 μm. (J–P) Representative blot images and quantitative analysis of phosphorylated and total NFκB P65, Nrf2, catalase, SOD3, SOD2 and SOD1. *P<0.05, **P<0.01, ***P<0.001 vs NG. #P<0.05, ##P<0.01, ###P<0.001 vs HG. n=4.

Article Snippet: Primary antibodies against catalase, SOD1, SOD2 and SOD3 were procured from Boster Biological Technology (Wuhan, China).

Techniques: Fluorescence, Staining

Journal: Virology

Article Title: Myeloid-Derived Suppressor Cells in Murine AIDS Inhibit B-Cell Responses in Part via Soluble Mediators including Reactive Oxygen and Nitrogen Species, and TGF-β

doi: 10.1016/j.virol.2016.08.031

Figure Lengend Snippet: Suppression of B-cell proliferation was assessed using supernates from suppressive cultures of MDSCs and B cells as in Figure 3. Supernates were treated with increasing doses of: (A) superoxide dismutase (SOD), (C) uric acid (UA), and (D) MnTBAP. Higher doses than those shown were tested, but did not show additional blockade and began to induce toxicity. (B) Superoxide levels in supernates were tested using the XTT assay; XO represents the positive control. Data shown are from representative experiments, these patterns of results were representative of at least 3 experiments for each panel. Significance levels:*, p<0.05; ***, p<0.001. p-values are versus untreated.

Article Snippet: Antioxidant Assays To test for involvement of reactive oxygen species, supernates were treated with catalase (MP Biomedicals, Santa Ana, CA), superoxide dismutase (SOD, MP Biomedicals, Santa Ana, CA), carboxy-PTIO (Sigma-Aldrich, St. Louis, MO), uric acid (UA, Pointe Scientific, Canton, MI), or MnTBAP (Enzo Life Sciences, Farmingdale, NY) at indicated concentrations to neutralize/scavenge hydrogen peroxide, superoxide, nitric oxide, or peroxynitrite, respectively.

Techniques: XTT Assay, Positive Control

Journal: Virology

Article Title: Myeloid-Derived Suppressor Cells in Murine AIDS Inhibit B-Cell Responses in Part via Soluble Mediators including Reactive Oxygen and Nitrogen Species, and TGF-β

doi: 10.1016/j.virol.2016.08.031

Figure Lengend Snippet: (A) Suppression of B-cell proliferation by MDSCs from LP-BM5-infected wild-type or VISTA/iNOS double knockout MDSCs. (B) Supernate transfer suppression assays (as in Fig. 3) left untreated or treated with superoxide dismutase (SOD) showed that soluble suppression by double knockout MDSCs accounted for about 2/3rds of total suppression, as opposed to with WT M-MDSCs, where soluble mediators account for approximately 55% the suppression (see Fig. 3a). Data shown are from a representative experiments. A similar pattern of results was observed in at least 3 experiments for each panel. Significance levels: **, p<0.01; NS, not significant.

Article Snippet: Antioxidant Assays To test for involvement of reactive oxygen species, supernates were treated with catalase (MP Biomedicals, Santa Ana, CA), superoxide dismutase (SOD, MP Biomedicals, Santa Ana, CA), carboxy-PTIO (Sigma-Aldrich, St. Louis, MO), uric acid (UA, Pointe Scientific, Canton, MI), or MnTBAP (Enzo Life Sciences, Farmingdale, NY) at indicated concentrations to neutralize/scavenge hydrogen peroxide, superoxide, nitric oxide, or peroxynitrite, respectively.

Techniques: Infection, Double Knockout

Journal: Redox Biology

Article Title: The loss of pancreatic islet NADPH oxidase (NOX)2 improves islet transplantation

doi: 10.1016/j.redox.2022.102419

Figure Lengend Snippet: The loss of NOX2 increases the expression of insulin and antioxidative proteins in hypoxic islets. ( A ) Representative Western blot analysis of (from top to bottom) Nrf2, β-actin, HO-1, SOD2, SOD1 and insulin from whole cell extracts of isolated hypoxic WT and Nox2 −/− islets. ( B ) Quantitative analysis of insulin expression (Fold change) (n = 5 each). Mean ± SEM. *P < 0.05 vs. WT. ( C ) Quantitative analysis of SOD1 expression (Fold change) (n = 3 each). Mean ± SEM. *P < 0.05 vs. WT. ( D ) Quantitative analysis of SOD2 expression (Fold change) (n = 3 each). Mean ± SEM. *P < 0.05 vs. WT. ( E ) Quantitative analysis of HO-1 expression (Fold change) (n = 5 each). Mean ± SEM. *P < 0.05 vs. WT. ( F ) Quantitative analysis of Nrf2 expression (Fold change) (n = 5 each). Mean ± SEM. *P < 0.05 vs. WT.

Article Snippet: The anti-SOD1 (TA321133) and anti-SOD2 (TA321189) antibodies were purchased from OriGene (Rockville, USA).

Techniques: Expressing, Western Blot, Isolation

Journal: Pharmaceuticals

Article Title: Dapagliflozin/Hesperidin Combination Mitigates Lipopolysaccharide-Induced Alzheimer’s Disease in Rats

doi: 10.3390/ph16101370

Figure Lengend Snippet: Effect of dapagliflozin and/or hesperidin on ( A ) malondialdehyde, ( B ) total antioxidant capacity, ( C ) catalase, ( D ) superoxide dismutase, ( E ) paraoxonase-1, and ( F ) Nrf2 content of the hippocampal tissues of rats injected with lipopolysaccharide (LPS) (Mean ± SD). a p -value < 0.05 versus the control group; b p -value < 0.05 versus LPS-injected group; c p -value < 0.05 versus LPS rats treated with dapagliflozin; d p -value < 0.05 versus LPS rats treated with hesperidin. LPS (lipopolysaccharide), DMSO (dimethyl sulfoxide), DPF (dapagliflozin), and HSP (hesperidin).

Article Snippet: The levels of catalase (CAT) and superoxide dismutase (SOD) were measured in the hippocampal tissues using kits purchased from Shanghai Korain Biotech Co., Shanghai, China (code number E0869Ra and E1444Ra, respectively).

Techniques: Injection, Control