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

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

Journal: Comparative Immunology Reports

Article Title: Dietary lacto-sacc stimulates the immune response of gravid mud crab (Scylla olivacea)

doi: 10.1016/j.cirep.2024.200156

Figure Lengend Snippet: Fig. 3. (A) Phenoloxidase (PO) and prophenoloxidase (proPO) activity asay; (B) Superoxide dismutase (SOD) activity assay.

Article Snippet: Analysis of superoxide dismutase activity assay The SOD activity of gravid mud crabs was tested using the Ghosh et al. [42] method and the Creative BioMart, Inc., USA procedure (EC 1.15.1.1) [42].

Techniques: Activity Assay

Journal: eLife

Article Title: CPT1A mediates radiation sensitivity in colorectal cancer

doi: 10.7554/elife.97827

Figure Lengend Snippet: Figure 6. CPT1A increases the transcription and protein of reactive oxygen species (ROS) scavenge-related genes by regulating the transcription factor activity of FOXM1. (A) The protein level of FOXM1, CPT1A, catalase (CAT), SOD1, SOD2, SOD3 after knockout and overexpression of CPT1A. (B) The mRNA level of FOXM1, CAT, SOD1, SOD2, SOD3 after knockout and overexpression of CPT1A. (C) Venn diagram showing the potential transcription factor of SOD1, SOD2, and CAT. (D) The protein level of FOXM1, CPT1A, CAT, SOD1, SOD2 after overexpression of FOXM1 in HCT116-CPT1AKO cells.

Article Snippet: DOI: https://doi.org/10.7554/eLife.97827 14 of 21 Reagent type (species) or resource Designation Source or reference Identifiers Additional information Antibody SOD1 Rabbit Polyclonal antibody Proteintech (China) 10269- 1- AP Antibody SOD2 Rabbit Polyclonal antibody Proteintech (China) 24127- 1- AP Antibody SOD3 Rabbit Polyclonal antibody Proteintech (China) 14316- 1- AP Antibody Catalase Rabbit Polyclonal antibody Proteintech (China) 21260- 1- AP Antibody FOXM1 Rabbit Polyclonal antibody Proteintech (China) 13147- 1- AP Antibody Anti- rabbit IgG, HRP- linked Antibody Cell Signaling Technology (USA) 7074 Chemical compound, drug TRIzol TakaraBio (Japan) 9108 Commercial assay or kit Evo M- MLV RT Mix Kit Accurate Biotechnology (China) AG11728 Commercial assay or kit SYBR Green Premix Pro Taq HS qPCR Kit Accurate Biotechnology (China) AG11701 Commercial assay or kit Protein extraction kit KeyGen BioTech (China) KGP113- SDS Commercial assay or kit Comet assay kit KeyGen BioTech (China) KGA240 Chemical compound, drug PI Beyotime (China) ST511 Chemical compound, drug DAPI Beyotime (China) C1002 Commercial assay or kit ROS detection kit Beyotime (China) S0033S Commercial assay or kit GSH detection kit Solarbio (China) BC1175 Commercial assay or kit GSSG detection kit Solarbio (China) BC1180 Commercial assay or kit SOD enzyme activity kit Solarbio (China) BC0170 Commercial assay or kit CAT enzyme activity kit Solarbio (China) BC0200 Commercial assay or kit POD enzyme activity kit Solarbio (China) BC0090 Continued

Techniques: Activity Assay, Knock-Out, Over Expression