Journal: Revista Portuguesa de Cardiologia (English Edition)

Article Title: Dexmedetomidine attenuates H2O2-induced apoptosis of rat cardiomyocytes independently of antioxidant enzyme expression

doi: 10.1016/j.repce.2020.07.015

Figure Lengend Snippet: Figure 3 Dexmedetomidine suppresses mRNA expression of superoxide dismutase 2 (SOD2), glutathione peroxidase 4 (GPX4), glutaredoxin

Article Snippet: The primary antibodies used were as follows: SOD2 (1:1000; Cell Signaling Technology, MA, USA); GPX4 (1:1000; Cell Signaling Technology, MA, USA); Grx1 (1:2000; Cell Signaling Technology, MA, USA); catalase (1:2000; Cell Signaling Technology, MA, USA) and beta-actin (1:2000; Cell Signaling Technology, MA, USA).

Techniques: Expressing

Journal: Revista Portuguesa de Cardiologia (English Edition)

Article Title: Dexmedetomidine attenuates H2O2-induced apoptosis of rat cardiomyocytes independently of antioxidant enzyme expression

doi: 10.1016/j.repce.2020.07.015

Figure Lengend Snippet: Figure 4 Dexmedetomidine suppresses protein expression of superoxide dismutase 2 (SOD2), glutathione peroxidase 4 (GPX4), glutaredoxin 1 (Grx1), and catalase in cardiomyocytes. Protein expression of antioxidant enzymes was detected by Western blot anal- ysis. Relative protein expression levels were normalized to beta-actin. Statistical significance was determined using one-way analysis of

Article Snippet: The primary antibodies used were as follows: SOD2 (1:1000; Cell Signaling Technology, MA, USA); GPX4 (1:1000; Cell Signaling Technology, MA, USA); Grx1 (1:2000; Cell Signaling Technology, MA, USA); catalase (1:2000; Cell Signaling Technology, MA, USA) and beta-actin (1:2000; Cell Signaling Technology, MA, USA).

Techniques: Expressing, Western Blot

Journal: PLoS ONE

Article Title: Protective effects and functional mechanisms of Lactobacillus gasseri SBT2055 against oxidative stress

doi: 10.1371/journal.pone.0177106

Figure Lengend Snippet: (A-D) MEF cells were cultured with or without LG2055 for 24 h. (A, D) The total cell lysates were analyzed by western blotting to compare the Nrf2 or SOD2 protein levels. The relative expression level of Nrf2 or SOD2 normalized by β-actin protein expression was quantitated. (B) The cells were fractionated into cytosolic and nuclear fractions using the Focus SubCell kit. Each fraction was analyzed by western blotting to compare the Nrf2 protein levels. Lamin B1 and Hsp90 were detected as loading controls. (C) The mRNA expression levels of cytoprotective genes were determined by quantitative real time-PCR analysis. (A, C, D) Each experiment was performed in triplicate; the data are shown as the means ± SD. ** p <0.01 and *** p <0.001 according to the Student’s t -test.

Article Snippet: After transfer, the membranes were blocked with 5% bovine serum albumin or 5% skimmed milk in Tris-buffered saline containing 0.1% Tween 20 (TBS-T) for 1 h at room temperature and incubated overnight at 4°C with primary antibody as follows: Nrf2 (D1Z9C), Phospho-JNK Thr183/Tyr185 (81E11), JNK, Phospho-c-Jun Ser73 (D47G9), c-Jun (60A8), SOD2 (D3X8F) (all from Cell Signaling Technology, Danvers, MA), Lamin B1 (EPR8985(B), Abcam, Cambridge, UK), Hsp90 (68, BD Biosciences), and β-actin (13E5, Cell Signaling).

Techniques: Cell Culture, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Oxidative Medicine and Cellular Longevity

Article Title: In Situ Evaluation of Oxidative Stress in Rat Fatty Liver Induced by a Methionine- and Choline-Deficient Diet

doi: 10.1155/2016/9307064

Figure Lengend Snippet: Liver of rats submitted to control diet for 2 weeks. (a) Representative photomicrographs of PAS reaction for glycogen, abundant in all hepatocytes. (b) Nile Red-induced fluorescence of small neutral lipid droplets in the perisinusoidal cytoplasm of hepatocytes. (c) Diaminobenzidine-Mn 2+ -Co 2+ reaction for Reactive Oxygen Species (ROS); intense reaction in periportal hepatocytes and occasional Kupffer cells in the midzone or pericentral regions. (d) Immunoreaction against dinitrophenyl (DNP) groups for demonstrating carbonyl groups derivatized with dinitrophenyl hydrazine (DNPH); these are present in perisinusoidal and canalicular membrane domains of hepatocytes. (e) Immunoreaction for visualizing the expression of Mn-dependent Superoxide Dismutase 2 (SOD2); moderately intense staining in the cytoplasm of periportal and pericentral hepatocytes. P: branch of portal vein; CL: branch of centrolobular vein. Scale bar: 50 μ m (insets in (d) and (e) represent image details at higher magnification).

Article Snippet: Endogenous peroxidase activity blocking was performed with 10% methanol-3% H 2 O 2 in PBS for 20 min. After washing twice for 5 min in PBS, nonspecific site blocking was performed with 2.5% normal horse serum in PBS for 1 h. The sections were then incubated with a primary rabbit polyclonal antibody against SOD2 (NB100-1992; Novus Biologicals, Littleton, CO 80120, USA) diluted 1 : 300 in 2.5% normal horse serum at room temperature for 1 h. After two further 5 min washing instances in PBS, the sections were incubated with ImmPRESS HRP Universal Antibody (anti-mouse Ig/anti-rabbit Ig, peroxidase) Polymer Detection Kit (MP-7500; Vector Laboratories, Burlingame, CA 94010, USA) for 30 min at room temperature.

Techniques: Control, Fluorescence, Membrane, Expressing, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: In Situ Evaluation of Oxidative Stress in Rat Fatty Liver Induced by a Methionine- and Choline-Deficient Diet

doi: 10.1155/2016/9307064

Figure Lengend Snippet: Expression of Mn-dependent Superoxide Dismutase 2 (SOD2) in the liver of rats fed with the MCD diet for 1–4 weeks. Representative photomicrographs. Immunoreactivity is concentrated in the thin rim of cytoplasm surrounding the large lipid droplets. P: branch of portal vein; CL: branch of centrolobular vein. Scale bar: 50 μ m.

Article Snippet: Endogenous peroxidase activity blocking was performed with 10% methanol-3% H 2 O 2 in PBS for 20 min. After washing twice for 5 min in PBS, nonspecific site blocking was performed with 2.5% normal horse serum in PBS for 1 h. The sections were then incubated with a primary rabbit polyclonal antibody against SOD2 (NB100-1992; Novus Biologicals, Littleton, CO 80120, USA) diluted 1 : 300 in 2.5% normal horse serum at room temperature for 1 h. After two further 5 min washing instances in PBS, the sections were incubated with ImmPRESS HRP Universal Antibody (anti-mouse Ig/anti-rabbit Ig, peroxidase) Polymer Detection Kit (MP-7500; Vector Laboratories, Burlingame, CA 94010, USA) for 30 min at room temperature.

Techniques: Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: In Situ Evaluation of Oxidative Stress in Rat Fatty Liver Induced by a Methionine- and Choline-Deficient Diet

doi: 10.1155/2016/9307064

Figure Lengend Snippet: Evaluation of SOD2 expression in livers of the rats fed with control or MCD diet for 1–4 weeks. (a) Comparison of SOD2 expression in the liver of rats fed with control and MCD diet for 1–4 weeks. Histograms representing the ratio between optical density (OD) values of homogenates of the liver of rats fed with the MCD diet for 1 to 4 weeks and the OD of control livers (b).

Article Snippet: Endogenous peroxidase activity blocking was performed with 10% methanol-3% H 2 O 2 in PBS for 20 min. After washing twice for 5 min in PBS, nonspecific site blocking was performed with 2.5% normal horse serum in PBS for 1 h. The sections were then incubated with a primary rabbit polyclonal antibody against SOD2 (NB100-1992; Novus Biologicals, Littleton, CO 80120, USA) diluted 1 : 300 in 2.5% normal horse serum at room temperature for 1 h. After two further 5 min washing instances in PBS, the sections were incubated with ImmPRESS HRP Universal Antibody (anti-mouse Ig/anti-rabbit Ig, peroxidase) Polymer Detection Kit (MP-7500; Vector Laboratories, Burlingame, CA 94010, USA) for 30 min at room temperature.

Techniques: Expressing, Control, Comparison