Journal: Journal of animal science and biotechnology

Article Title: Increased maternal consumption of methionine as its hydroxyl analog improves placental angiogenesis and antioxidative capacity in sows.

doi: 10.1186/s40104-025-01159-z

Figure Lengend Snippet: Fig. 7 Increased consumption of methionine as OHMet improved angiogenesis and antioxidative capacity in pTr cells. A The trophoblast cells were treated for 24 h with 20% serum from each group (n = 3). B Cell viability. C Reactive oxidative species level. D Schematic of the Met metabolism and list of genes directly involved and associated with this pathway. E The gene expression of transmethylation and trans-sulphuration. F The gene expression of anti-oxidation. G The gene expression of angiogenesis. E–G Duplicated independent experiments. H The protein expression of pTr, GAPDH as a loading control. TXN, thioredoxin; GSTT1, glutathione transferase T1; VEGF-A, vascular endothelial growth factor A. Significant difference among four treatments is marked with different superscripts (P < 0.05). Abbreviations: CON = basal diet; 1.5S-OHMet = basal diet + 1.5 g/kg OHMet; 3.0S-OHMet = basal diet + 3.0 g/kg OHMet; 3.0S-Met = basal diet + 3.0 g/kg Met

Article Snippet: Primary antibodies were thioredoxin (TXN, Zen BioScience, 1:1,000), HADHA (Zen BioScience, 1:1,000), NDUFA3 (Zen BioScience, 1:1,000), ENO3 (Zen BioScience, 1:1,000), GSTT1 (Proteintech, 1:1,000), GAPDH (Absin, 1:1,000), β-actin (Cell Signaling Technology, 1:1,000).

Techniques: Gene Expression, Expressing, Control

Journal: Journal of animal science and biotechnology

Article Title: Increased maternal consumption of methionine as its hydroxyl analog improves placental angiogenesis and antioxidative capacity in sows.

doi: 10.1186/s40104-025-01159-z

Figure Lengend Snippet: Fig. 8 Anti-proliferative and inhibited antioxidant capacity induced by homocysteine in pTr cells (n = 3). A Detection of LDH activity in pTr cells. B EdU-positive cells were measured using ImageJ software. C Cell proliferation was determined by the EdU assay, and images were taken under a fluorescence microscope (magnification 400 × , bar = 100 μm). D Cell migration was measured using ImageJ software. E The cell migration distance (bar = 500 μm) and images of ROS were taken using a DCFH-DA probe (magnification 400 × , bar = 500 μm). F Representative western blot results for protein levels of enolase 3 (ENO3) and thioredoxin (TXN), β-actin was used as a loading reference. G–J Detection of apoptosis in pTr cells. Annexin V and propidium iodide (PI) fluorescence was quantified using flow cytometry. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Primary antibodies were thioredoxin (TXN, Zen BioScience, 1:1,000), HADHA (Zen BioScience, 1:1,000), NDUFA3 (Zen BioScience, 1:1,000), ENO3 (Zen BioScience, 1:1,000), GSTT1 (Proteintech, 1:1,000), GAPDH (Absin, 1:1,000), β-actin (Cell Signaling Technology, 1:1,000).

Techniques: Activity Assay, Software, EdU Assay, Fluorescence, Microscopy, Migration, Western Blot, Flow Cytometry

Journal: Scientific Reports

Article Title: HLA-DR cancer cells expression correlates with T cell infiltration and is enriched in lung adenocarcinoma with indolent behavior

doi: 10.1038/s41598-021-93807-3

Figure Lengend Snippet: Mass cytometry antibody panel for lung adenocarcinoma.

Article Snippet: Thioredoxin , 146-Nd , Intracellular , 2G11/TRX , Fluidigm , 3146016B.

Techniques: Mass Cytometry

Journal: Oncology Letters

Article Title: Combined treatment with D-allose, docetaxel and radiation inhibits the tumor growth in an in vivo model of head and neck cancer

doi: 10.3892/ol.2018.7787

Figure Lengend Snippet: Change of mRNA expression after the D-allose and radiation treatment.

Article Snippet: For the Western blot analyses, proteins were separated on 10% SDS-PAGE gels, transferred to nitrocellulose membranes, blocked with 5% (w/v) non-fat dried milk in PBS, and incubated with anti-TXNIP (MBL, Nagoya, Japan), anti-TRX (MBL), and anti-GAPDH (14c10) antibodies (Cell Signaling Technology, Inc., Tokyo, Japan).

Techniques: Expressing, Irradiation

Journal: Oncology Letters

Article Title: Combined treatment with D-allose, docetaxel and radiation inhibits the tumor growth in an in vivo model of head and neck cancer

doi: 10.3892/ol.2018.7787

Figure Lengend Snippet: TNF-α, TXNIP and TRX expression after the treatment. Histological features of the skin region. Three weeks after the initial treatment, skin specimens were obtained and fixed with 4% PFA. To observe inflammatory changes, TNF-α antibody was used. (Aa) Normal, saline-treated skin was included as a control (bar, 50 µm). (Ab) Hyperkeratosis and epidermal thickening (arrows) were observed after radiotherapy, with strong TNF-α staining. (Ac) Weak TNF-α staining was observed with D-allose treatment (arrow). (Ad) Radiation-induced epidermal thickening and TNF-α staining were reduced with additional D-allose treatment (arrow). (B) Western blot analysis of TXNIP and TRX expression. Proteins were obtained from: 1, normal skin with saline treatment; 2, normal skin with D-allose treatment for 2 weeks; 3, tumor tissue with saline treatment; 4, tumor tissue with D-allose treatment for 2 weeks; and 5, tumor tissue with D-allose treatment for 3 weeks. TXNIP, thioredoxin interacting protein; TRX, thioredoxin; TNF-α, tumor necrosis factor-α.

Article Snippet: For the Western blot analyses, proteins were separated on 10% SDS-PAGE gels, transferred to nitrocellulose membranes, blocked with 5% (w/v) non-fat dried milk in PBS, and incubated with anti-TXNIP (MBL, Nagoya, Japan), anti-TRX (MBL), and anti-GAPDH (14c10) antibodies (Cell Signaling Technology, Inc., Tokyo, Japan).

Techniques: Expressing, Staining, Western Blot

Journal: BMC Biotechnology

Article Title: Preparation of an epitope-based recombinant diagnostic antigen specific to anti-phospholipase A 2 receptor 1 antibodies

doi: 10.1186/s12896-018-0448-8

Figure Lengend Snippet: SDS-PAGE analysis of small-scale expression a , large-scale expression and purification steps of R101 protein b , and western blot analysis of R101 protein* c , d , e . Lane 1, non-induced bacteria; lanes 2–6, expression levels of different single colonies; lane 7, non-induced bacteria; lane 8, total bacterial proteins after sonication; lane 9, flow-through fraction from affinity chromatography; lane 10, supernatant from the homogenate; lane 11, eluates washed with 300 mM imidazole in affinity chromatography; lane 12, eluates washed with 200 mM NaCl in DEAE chromatography; lane 13, non-induced bacteria; and lane 14, purified R101 protein * C. Mouse anti-Trx monoclonal antibody as primary antibody; *D. Mouse anti-His monoclonal antibody as primary antibody; and *E. Mouse anti-PLA 2 R monoclonal antibody as primary antibody. All of the secondary antibodies were goat anti-mouse IgG-HRP conjugates.

Article Snippet: After blocking non-specific antibody sites with 5% ( w / v ) Difco skim milk (Becton Dickinson, San Jose, MD, USA) in TBST (20 mM Tris-HCl, 500 mM NaCl, 0.05% Tween-20, pH 7.5), each of three nitrocellulose membranes were reacted with either mouse anti-Trx monoclonal antibody (1:2000; Merck Millipore), mouse anti-His monoclonal antibody (1:2000; Merck Millipore), or mouse anti-PLA 2 R monoclonal antibody (1:2000; Abcam).

Techniques: SDS Page, Expressing, Purification, Western Blot, Bacteria, Sonication, Affinity Chromatography, Chromatography

Journal: PLoS ONE

Article Title: Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System

doi: 10.1371/journal.pone.0007949

Figure Lengend Snippet: (A,B) Hela cells transiently transfected with Flag-Nm23-H1 were exposed to 5 mM H 2 O 2 for 1 h at 37°C and recovered for 6 h without H 2 O 2 . Endogenous Nm23-H1s were purified by immunoprecipitation using anti-Flag antibody. These immuno-complexes were separated on 12% SDS-PAGE under non-reducing condition and detected with silver-staining (A) and with western analysis using anti-Flag antibody (B). (C) Protein interacting with oxidized Nm23 as indicated arrow in was identified with MALDI-TOF MS as thioredoxin reductase (TrxR). (D) Thioredoxin reductase interacting with oxidized Nm23-H1 was confirmed by western analysis using anti-TrxR antibody.

Article Snippet: Monoclonal anti-Flag antibody (M2) was purchased from Sigma (St. Louis, MO, USA), monoclonal anti-Nm23-H1 antibody and anti-tubulin from Santa Cruz Biotechnology (Santa Cruz, CA, USA), horseradish peroxidase conjugated goat-anti-mouse IgG and goat anti rabbit IgG from Bio-Rad Laboratories (Richmond, CA, USA), polyclonal anti-TrxR1 and anti-Trx1 antibodies from Ab Frontier (Seoul, Korea).

Techniques: Transfection, Purification, Immunoprecipitation, SDS Page, Silver Staining, Western Blot

Journal: PLoS ONE

Article Title: Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System

doi: 10.1371/journal.pone.0007949

Figure Lengend Snippet: (A) Suggested scheme of oxido-reduction regulation of Nm23-H1 by TrxR-Trx-NADPH system. (B) TrxR assay was performed in solution containing 50 mM sodium phosphate, pH 7.0, 50 mM KCl, 1 mM EDTA, 100 µM Nm23-H1, 200 µM NADPH, 80 µM rat TrxR1, 50 µM Trx1, and monitored the absorbance change at 340 nm. Absorbance decreases by conversion of NADPH to NADP were measured as TR activity. It turns out that oxidized Nm23-H1 is a substrate of TrxR-Trx-NADPH system.

Article Snippet: Monoclonal anti-Flag antibody (M2) was purchased from Sigma (St. Louis, MO, USA), monoclonal anti-Nm23-H1 antibody and anti-tubulin from Santa Cruz Biotechnology (Santa Cruz, CA, USA), horseradish peroxidase conjugated goat-anti-mouse IgG and goat anti rabbit IgG from Bio-Rad Laboratories (Richmond, CA, USA), polyclonal anti-TrxR1 and anti-Trx1 antibodies from Ab Frontier (Seoul, Korea).

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: Multiple Functions of Nm23-H1 Are Regulated by Oxido-Reduction System

doi: 10.1371/journal.pone.0007949

Figure Lengend Snippet: (A,B) Recombinant Nm23-H1 proteins (100 µM) were oxidized and reincubated with 200 µM NADPH, 80 µM rat TrxR1 and various concentration of Trx (0, 10 and 50 µM) same as in . Enzymatic activities were measured on TLC plate (A) and quantitatively analyzed (B). (A) Same samples were separated on % SDS-PAGE under non-reducing condition.

Article Snippet: Monoclonal anti-Flag antibody (M2) was purchased from Sigma (St. Louis, MO, USA), monoclonal anti-Nm23-H1 antibody and anti-tubulin from Santa Cruz Biotechnology (Santa Cruz, CA, USA), horseradish peroxidase conjugated goat-anti-mouse IgG and goat anti rabbit IgG from Bio-Rad Laboratories (Richmond, CA, USA), polyclonal anti-TrxR1 and anti-Trx1 antibodies from Ab Frontier (Seoul, Korea).

Techniques: Recombinant, Concentration Assay, SDS Page