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: Redox Biology

Article Title: MG53 protein rejuvenates hUC-MSCs and facilitates their therapeutic effects in AD mice by activating Nrf2 signaling pathway

doi: 10.1016/j.redox.2022.102325

Figure Lengend Snippet: MG53 reduces oxidative stress in senescent hUC-MSCs by activating the Nrf2 signaling pathway. (A) ROS production measured by DCFH-DA kit. (B) MDA level. (C) SOD viability. (D) Protein expression of Nrf2 signaling pathway in each group. (E) Densitometric analysis of Nrf2 expression in the nucleus. (F) Densitometric analysis of Keap1, NQO1, SOD1 and Nrf2 in the cytosol. Data were presented as mean ± SEM. Data were presented as mean ± SEM. N = 3 per group. *: Compared with P5 group, P < 0.05; #: Compared with P10 group, P < 0.05; Δ: Compared with P15, P < 0.05.

Article Snippet: The primary antibodies were MG53 (1:1000), P16 (1:1000, C610021, Sangon, China), P21 (1:1000, D220091, Sangon, China), P53(1:1000, 10442-1-AP, Proteintech, China), Tau (1:1000, D121297, Sangon, China), p-Tau (Ser396) (1:1000, D155045, Sangon, China), p-Tau (Ser231) (1:1000, D155301, Sangon, China) and p-Tau (Ser235) (1:1000, D155045, Sangon, China), Nrf2 (1:1000, WL02135, Wanlei, China), SOD1 (1:1000, WL01846, Wanlei, China), NQO1 (1:1000, WL04860, Wanlei, China) and Keap-1 (1:1000, WL03285, Wanlei, China), pCNA (1:1000, 10205-2-AP, Proteintech, China), Sirt1 (1:2000, 13161-1-AP, Proteintech, China), β-actin (1:2000, 20536-1-AP, Proteintech, China) and Histone-3 (1:2000, WL0984a, Wanlei, China). β-Actin and Histone-3 were used as internal reference of protein expression in the cytosol and nucleus, respectively.

Techniques: Expressing

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: 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