Journal: Advanced Science

Article Title: Identification of PRDX5 as A Target for The Treatment of Castration‐Resistant Prostate Cancer

doi: 10.1002/advs.202304939

Figure Lengend Snippet: Upregulation of the thioredoxin/peroxiredoxin pathway in DTP cells A) LNCaP epiDTP or enzDTP cells were treated for 48 h with EPI001 (EPI 50 µM) or enzalutamide (ENZ 60 µM), respectively, plus each ninety‐three drugs (10 µM) identified in Figure . Experiments identified 9 drugs that reduced cell survival below 70%. Each data point represents the average of three independent experiments and is expressed as a percentage of viable cells relative to untreated controls as measured by CCK‐8 assay. Standard deviations are less than 5%. B) The highest concentration that does not affect LNCaP cell viability (IC 0 ) was determined for the above 9 drugs. Subsequently, LNCaP epiDTP or enzDTP cells were treated with each drug (IC 0 ) with or without EPI001 (EPI 50 µM) or enzalutamide (ENZ 60 µM) for 48 h. Additive or synergistic effects were seen for all nine drugs. Each data point represents the average of three independent experiments and is expressed as a percentage of viable cells relative to untreated controls as measured by CCK‐8 assay. Standard deviations are less than 5%. C) Combination indexes (CI) of nine drugs were measured. A CI of less than, equal to, and more than 1 indicates synergy, additivity, and antagonism, respectively. D) 22Rv1 cells that are resistant to ENZ were treated for 48 h with ENZ (60 µM), each of the 9 drugs identified above, or in combinations. Results indicated that all 9 drugs either alone or in combination with ENZ could reduce cell survival. Each data point represents the average of three independent experiments and is expressed as a percentage of viable cells relative to untreated controls as measured by CCK‐8 assay. Standard deviations are less than 5%. E) Diagram portrays the thioredoxin/peroxiredoxin pathway and up‐regulation of key proteins (red color) in DTP cells. F: Confirmation of proteomics results (TXNRD1, TXNRD2, TXNRD3, TXN2, and PRDX5) by Western blotting. GAPDH protein is used as the loading control. Data are expressed as mean ± std of triplicates. One‐way ANOVA with the Turkey test was performed. p <0.05 was considered significant and indicated by different letters.

Article Snippet: PRDX5 (67599‐1‐Ig; 1:2000), CDKN1A (p21) (60214‐1‐Ig; 1:1000), Cyclin E1 (CCNE1) (11554‐1‐AP; 1:1000), Cyclin B1 (CCNB1) (67686‐1‐Ig; 1:5000), CDC6 (11640‐1‐AP; 1:2000), CDC2 (CDK1) (19532‐1‐AP; 1:1000), TXNRD1 (67728‐1‐Ig; 1:10 000), TXNRD2 (16360‐1‐AP; 1:3000), TXNRD3 (19517‐1‐AP; 1:2000), TXN2 (13089‐1‐AP; 1:1000), NSE (66150‐1‐Ig; 1:10 000), CgA (60135‐2‐Ig; 1:3000), CgB (14968‐1‐AP; 1:2000), PTHrP (29115‐1‐AP; 1:800), AMACR (15918‐1‐AP; 1:500), GAPDH (60004‐1‐Ig; 1:50 000), β‐ACTIN (66009‐1‐Ig; 1:10 000) antibodies were purchased from Proteintech Group (IL, US). β‐TUBLIN (86 298; 1:10 000) antibody was purchased from Cell Signaling Technology (MA, US).

Techniques: CCK-8 Assay, Concentration Assay, Western Blot, 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. 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: RNA

Article Title: Anti-Argonaute RIP-Chip shows that miRNA transfections alter global patterns of mRNA recruitment to microribonucleoprotein complexes

doi: 10.1261/rna.1905910

Figure Lengend Snippet: Western blot analysis after miRNA transfections helped validate RIP-Chip results. H4 cells were first transfected with full-length cDNA plasmids: pCMV6-LDLR, pCMV6-TXNIP, and pCMV6-DBI for 24 h, and were then transfected with 25 nM of miR-107, miR-124, miR-128, miR-320 precursors, or a negative control miRNA for additional 48 h. Cells were harvested, and Western blot analyses were performed using antibodies against each expressing proteins. Anti-β-actin antibody was used to probe each blot to monitor the total protein loadings. The three miR-128 targeted proteins shown were effectively knocked down by miR-128 transfection. LDLR was also strongly reduced by miR-124 transfection. RIP-Chip did not identify LDLR as miR-124 target, however, microarray analysis of total cell RNA showed that LDLR mRNA was significant decreased upon miR-124 transfection.

Article Snippet: Full-length cDNA (all UTR-containing) cloned in pCMV6-XL5 plasmid vectors were obtained that express human low density lipoprotein receptor (LDLR, {"type":"entrez-nucleotide","attrs":{"text":"NM_000527.2","term_id":"8051613","term_text":"NM_000527.2"}} NM_000527.2 ), thioredoxin interacting protein (TXNIP, {"type":"entrez-nucleotide","attrs":{"text":"NM_006472","term_id":"928192547","term_text":"NM_006472"}} NM_006472 ), and diazepam binding inhibitor (DBI, {"type":"entrez-nucleotide","attrs":{"text":"NM_020548.4","term_id":"54262129","term_text":"NM_020548.4"}} NM_020548.4 ) (OriGene Technologies, Inc.).

Techniques: Western Blot, Transfection, RNA Immunoprecipitation - Chromatin Immunoprecipitation, Negative Control, Expressing, Microarray