Journal: Toxins

Article Title: Inhibition of Aflatoxin Production by Paraquat and External Superoxide Dismutase in Aspergillus flavus

doi: 10.3390/toxins11020107

Figure Lengend Snippet: Determination of SOD activity and SOD protein abundance. ( a – c ) A. flavus was cultured for 48 h. SOD activity of fungal cell extracts ( a ) and culture supernatants ( b ) was determined, and fungal cell extracts were subjected to western blotting using anti-human SOD1 antibody ( c ). The density of the bands indicated by the arrow was quantified using Image J. ( d – f ) After A. flavus was cultured for 24 h, mycelia were collected, washed with distilled water, transferred to fresh medium, and incubated for another 24 h. Then SOD activity in fungal cell extracts ( d ) and culture supernatant ( e ) was determined. Fungal cell extracts were subjected to western blotting using anti-human SOD1 antibody, and the density of the bands indicated by the arrow was quantified ( f ). Data are presented as means and standard deviations from three biological replicates. Asterisks indicate significant differences (* P < 0.05, ** P < 0.01 vs. control group, Dunnett test).

Article Snippet: The membrane was immuno-blotted using anti-SOD1 antibody (SPC-115C; StressMarq Biosciences, British Columbia, Canada) followed by goat anti-rabbit IgG (H+L) poly-horseradish peroxidase antibody (32260; Thermo Fisher Scientific).

Techniques: Activity Assay, Cell Culture, Western Blot, Incubation, Control

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: The Journal of physiology

Article Title: Initiation of 3,3-dimethyl-1-butanol at midlife prevents endothelial dysfunction and attenuates in vivo aortic stiffening with ageing in mice

doi: 10.1113/JP283581

Figure Lengend Snippet: A, carotid artery endothelium-independent dilatation (EID) in response to increasing doses of the NO donor sodium nitroprusside (SNP); and B, EID area under the curve (AUC) (both n = 17–21 per group). C, EDD to acetylcholine (ACh) in the presence of the NO synthase inhibitor NG-nitro-l-arginine methyl ester (l-NAME); and D, NO-mediated dilatation, as assessed by the difference in peak EDD in the absence vs. presence of l-NAME (both n = 9–13 per group). E, superoxide production, measured in 1 mm aorta rings by electron paramagnetic resonance (EPR) spectroscopy, with representative tracings shown above (n = 11–22 per group). Aortic abundance of superoxide dismutase (SOD) 1 (F) and 2 (G) normalized to GAPDH with representative Western blot images generated from WES electropherograms shown above. H, peak EDD to ACh in the absence vs. presence of the superoxide dismutase mimetic 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPOL; n = 8–15 per group). Data are mean ± SD. Data are combined for mice aged 24 and 27 months. Statistics are two-way mixed (group dose) ANOVA with Tukey’s post hoc test (A and C), one-way ANOVA with Tukey’s post hoc test (B, D and E),×and two-way mixed (group × ACh alone or ACh + TEMPOL) with Šídák’s post hoc test (F). *P < 0.05 control vs. young 5 month reference group within dose/condition. †P < 0.05 control vs. DMB within dose/condition. ˆP < 0.01 DMB vs. young 5 month reference group within dose. Abbreviations: Y, young; OC, old control; OD, old DMB.

Article Snippet: Primary antibodies were anti-SOD1/Cu-Zn SOD (1:500; R&D, Minneapolis, MN, USA; Cat# AF3787), anti-SOD2/Mn-SOD (1:50; R&D; Cat# AF3419) and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:200, Cell Signaling Technology, Danvers, MA, USA; Cat # 14C10).

Techniques: Electron Paramagnetic Resonance, Spectroscopy, Western Blot, Generated

Journal: Viruses

Article Title: Extracellular Vesicles from Human Papilloma Virus-Infected Cervical Cancer Cells Enhance HIV-1 Replication in Differentiated U1 Cell Line

doi: 10.3390/v12020239

Figure Lengend Snippet: Caski cell culture supernatant (CCS) alters antioxidant enzyme (AOE) expression in differentiated U1 cell line. Differentiated U1 cell line was treated with 250 µL of CCS every 24 h for 4 days. Expressions of SOD1, SOD2, catalase, and PRDX6 were monitored at mRNA ( A–D ) and protein ( E–H ) levels in the treated cells. Data were obtained from the mean of at least three independent experiments with the error bars representing the standard error of the mean.

Article Snippet: We used the following primary antibodies: GAPDH Rabbit Mab, 1:2000 dilution (Cell Signaling Technology, Danvers, MA, USA), catalogue #2118; CYP1A1 rabbit Mab, 1:200 dilution (Proteintech Group, Inc., Rosemont, IL, USA), catalogue #13241-1-AP; CYP1B1 Rabbit Mab, 1:500 dilution (Santa Cruz Biotechnology, Dallas, TX, USA), catalogue #sc-32882; CYP2A6 Mouse Mab, 1:200 dilution (Abcam, Cambridge, MA, USA), catalogue #ab3570; SOD1 Mouse Mab, 1:1500 dilution, catalog #sc-101523; SOD2 Mouse Mab, 1:500 dilution, catalogue #sc-133254; Catalase Mouse Mab, 1:1200 dilution (Santa Cruz Biotechnology Inc., Dallas, TX, USA), catalog #21260-1-AP; PRDX6 Rabbit Mab, 1:500 dilution (LifeSpan Biosciences, Inc., Seattle, WA, USA), catalog #LS-C162131; CD63, Rabbit Pab, 1:200 dilution (Proteintech Group, Rosemont, IL, USA), catalog #25682-1-AP.

Techniques: Cell Culture, Expressing

Journal: Viruses

Article Title: Extracellular Vesicles from Human Papilloma Virus-Infected Cervical Cancer Cells Enhance HIV-1 Replication in Differentiated U1 Cell Line

doi: 10.3390/v12020239

Figure Lengend Snippet: Differentiated U1 cell line uptake CCS-derived EVs (CCS-EVs) that contain oxidative stress factors. ( A ) The proteins obtained from CCS-derived EVs (CCS-EVs) were examined for the expression of EVs markers, CD63 and CD81, CYPs (1A1, 1B1, 2A6) and AOEs (SOD1, SOD2, and catalase) and HPV 16 oncoproteins (E6 and E7). ( B ) To monitor the EVs uptake by U1 cells, we labeled the CCS-EVs with GFP (Green Fluorescent Protein) and treated the labeled EVs to U1 cells. After 6 h of incubation, we monitored the fluorescent intensity of GFP under fluorescent microscope. ( C ) The GFP fluorescence was further quantified using flow cytometry.

Article Snippet: We used the following primary antibodies: GAPDH Rabbit Mab, 1:2000 dilution (Cell Signaling Technology, Danvers, MA, USA), catalogue #2118; CYP1A1 rabbit Mab, 1:200 dilution (Proteintech Group, Inc., Rosemont, IL, USA), catalogue #13241-1-AP; CYP1B1 Rabbit Mab, 1:500 dilution (Santa Cruz Biotechnology, Dallas, TX, USA), catalogue #sc-32882; CYP2A6 Mouse Mab, 1:200 dilution (Abcam, Cambridge, MA, USA), catalogue #ab3570; SOD1 Mouse Mab, 1:1500 dilution, catalog #sc-101523; SOD2 Mouse Mab, 1:500 dilution, catalogue #sc-133254; Catalase Mouse Mab, 1:1200 dilution (Santa Cruz Biotechnology Inc., Dallas, TX, USA), catalog #21260-1-AP; PRDX6 Rabbit Mab, 1:500 dilution (LifeSpan Biosciences, Inc., Seattle, WA, USA), catalog #LS-C162131; CD63, Rabbit Pab, 1:200 dilution (Proteintech Group, Rosemont, IL, USA), catalog #25682-1-AP.

Techniques: Derivative Assay, Expressing, Labeling, Incubation, Microscopy, Fluorescence, Flow Cytometry

Journal: International Journal of Environmental Research and Public Health

Article Title: Alcohol-Induced Blood-Brain Barrier Impairment: An In Vitro Study

doi: 10.3390/ijerph18052683

Figure Lengend Snippet: Primary antibody for Western blotting analysis.

Article Snippet: SOD1 SOD2 , 1:5000 , GeneTex, Prodotti Gianni, Milan, Italy , rabbit.

Techniques: Western Blot

Journal: International Journal of Environmental Research and Public Health

Article Title: Alcohol-Induced Blood-Brain Barrier Impairment: An In Vitro Study

doi: 10.3390/ijerph18052683

Figure Lengend Snippet: SOD1 and SOD2 expression levels on RBE4 cells after EtOH treatment. Western blotting analysis and quantification of SOD1 (panel A ) and SOD2 (panel B ) protein expression levels during EtOH (50, 75 and 100 mM) treatments at 2 and 4 h. Values are expressed in percentage of control (untreated cells) as the mean ± S.E.M. of three independent experiments in triplicate. * p < 0.05 vs. control.

Article Snippet: SOD1 SOD2 , 1:5000 , GeneTex, Prodotti Gianni, Milan, Italy , rabbit.

Techniques: Expressing, Western Blot, Control