Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: Women with hsCRP have high levels of GDF15. (a, b) Serum GDF15 levels were quantified by ELISA from HANDLS participants with either low- (<3 mg/L), mid- (>3–20 mg/L), or high hsCRP (>20 mg/L) levels ( n = 39/group). The ELISA assay was performed according to manufacturer's instructions and was repeated in 2 independent experiments. (c) RNA was isolated from PBMCs from HANDLS participants with either low- (<3 mg/L) or high hsCRP (>20 mg/L) levels ( n = 15/group). GDF15 mRNA was quantified by RT-qPCR and normalized to HPRT1 and UBC levels. The histograms represent the mean + SEM from three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 by Student's t -test.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Quantitative RT-PCR

Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: CRP upregulates GDF15 expression. (a) 18 h after CRP treatment with the indicated doses, GDF15 expression in HAECs was analyzed by immunoblotting with anti-GDF15 antibodies. β -Actin was used as a loading control. (b) After CRP treatment for the indicated time points, conditioned media was collected and GDF15 secreted levels were analyzed by ELISA. GDF15 levels were normalized to the 0 h time point for each experiment. The mean of three independent experiments is shown. (c and d) 18 h after CRP (25 μ g/mL) treatment, HAECs were lysed and levels of GDF15 mRNA or protein were quantified by RT-qPCR analysis (c) and western blot analysis (d). (e) HAECs were transfected with pCMV6-control or pCMV6-CRP plasmid for 48 h. Total RNA was isolated, and mRNA levels were quantified using RT-qPCR and normalized to GAPDH . (f) Total cell lysates from the indicated transfected HAECs were analyzed by Western blotting. β -Actin was used as a loading control. The histograms represent the mean + SEM from three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 by Student's t -test.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Expressing, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Transfection, Plasmid Preparation, Isolation

Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: CRP promotes GDF15 transcription. (a) Schematic of GDF15 promotor dual-luciferase constructs. Two p53 binding sites are indicated. (b) The indicated plasmids (1 μ g) were cotransfected with 0.1 μ g of TK-Renilla reporter plasmid in HeLa cells, and 24 h later, the cells were treated with CRP. After 18 h, the promoter activities were measured by luciferase activity. Transfection efficiency for luciferase activity was normalized to the Renilla luciferase activity. The results show the mean + SEM of three independent transfections. ∗∗ p < 0.01 by Student's t -test. (c) Schematic of p53 binding sites and primers used for ChIP assays in the GDF15 promoter. (d) ChIP assays were performed on HAECs transfected for 24 h and treated with or without CRP for 18 h. DNA immunoprecipitated by antibodies to p53 or immunoglobulin G IgG (control) was amplified by qPCR. Each qPCR reaction was performed in triplicate, and the histogram represents the average of three independent ChIP assays + SEM.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Luciferase, Construct, Binding Assay, Plasmid Preparation, Activity Assay, Transfection, Immunoprecipitation, Control, Amplification

Journal: Mediators of Inflammation

Article Title: CRP Stimulates GDF15 Expression in Endothelial Cells through p53

doi: 10.1155/2018/8278039

Figure Lengend Snippet: p53 knockdown inhibits CRP-induced GDF15 expression. HAECs were transfected with either Ctrl siRNA or p53 siRNA for 24 h and treated with or without CRP for 18 h. GDF15 mRNA levels were examined by RT-qPCR (a), and protein levels were analyzed by Western blot analysis (b). The histogram represents the mean + SEM from three independent experiments. ∗ p < 0.05 and ∗∗ p < 0.01 by Student's t -test.

Article Snippet: Human GDF15 Quantikine ELISA Kit (R&D Systems) was used according to the manufacturer's directions.

Techniques: Knockdown, Expressing, Transfection, Quantitative RT-PCR, Western Blot

Journal: Journal of Personalized Medicine

Article Title: Growth Differentiation Factor-15 as an Emerging Biomarker in Cardiology: Diagnostic and Prognostic Implications

doi: 10.3390/jpm16010016

Figure Lengend Snippet: Schematic representation of GDF-15 biosynthesis, maturation, and secretion. ( a ) GDF-15 is synthesized as pre-pro-GDF-15, consisting of a signal peptide (blue), a pro-domain (yellow), and a mature C-terminal domain (red). Removal of the signal peptide generates pro-GDF-15, which rapidly dimerizes; ( b ) In the Golgi apparatus, pro-GDF-15 dimer is cleaved by PCSK3, PCSK5, and PCSK6 at the RXXR site, producing mature GDF-15; ( c ) Mature GDF-15 homodimers are secreted through the classical secretory pathway and released into circulation, while in specific cells pro-GDF-15 dimers may remain bound to the extracellular matrix until further cleavage by MMP26 or PCSKs. GDF-15: Growth Differentiation Factor-15; PCSK: proprotein convertase subtilisin–kexin; MMP26: matrix metalloproteinase 26.

Article Snippet: Human GDF-15 ELISA Kit [ ] , Proteintech , ELISA , 300 , 1250–40,000.

Techniques: Synthesized

Journal: Journal of Personalized Medicine

Article Title: Growth Differentiation Factor-15 as an Emerging Biomarker in Cardiology: Diagnostic and Prognostic Implications

doi: 10.3390/jpm16010016

Figure Lengend Snippet: Molecular regulation of GDF-15. Inflammatory and stress-related mediators (IL-1β, TNF-α, IL-2, M-CSF) regulate GDF-15 expression by activating key transcription factors such as ATF4, CHOP, p53, EGR-1, and NF-κB, emphasizing the complex and tissue-dependent nature of this regulatory network. IL-1ß: interleukin-1ß; TNF-α: tumor necrosis factor-α, IL-2: interleukin-2; MCSF-1: macrophage colony-stimulating factor-1; EGR-1: early growth response transcription factor 1; NF-Kb: Nuclear Factor kappa B; CHOP: C/EBP homologous protein; ATF4: activating transcription factor 4; GDF-15: Growth Differentiation Factor-15.

Article Snippet: Human GDF-15 ELISA Kit [ ] , Proteintech , ELISA , 300 , 1250–40,000.

Techniques: Expressing

Journal: Journal of Personalized Medicine

Article Title: Growth Differentiation Factor-15 as an Emerging Biomarker in Cardiology: Diagnostic and Prognostic Implications

doi: 10.3390/jpm16010016

Figure Lengend Snippet: GDF-15–mediated mechanisms of cardiac protection. In cardiomyocytes, GDF-15 activates the Smad2/3 and Smad1/5/8 pathways to limit hypertrophy and apoptosis, enhances myocardial survival through the PI3K/PKB signaling cascade, and inhibits pro-apoptotic pathways involving JNK, Bad, and EGFR, thereby contributing to cardiac protection. GDF-15: Growth Differentiation Factor-15; EGFR: epidermal growth factor receptor; ERK: extracellular signal-regulated kinase; PKB: protein kinase B; NF-Kb: Nuclear Factor kappa B; JNK: c-Jun N-terminal kinase; CAS-3: Caspase-3; PI3K: phosphatidylinositol 3-kinase; BAD: Bcl-2-associated death promoter.

Article Snippet: Human GDF-15 ELISA Kit [ ] , Proteintech , ELISA , 300 , 1250–40,000.

Techniques:

Journal: Mediators of inflammation

Article Title: Potential Association of Gut Microbial Metabolism and Circulating mRNA Based on Multiomics Sequencing Analysis in Fetal Growth Restriction.

doi: 10.1155/2024/9986187

Figure Lengend Snippet: FIGURE 2: Validation of DEGs in circulation and placental tissues: (a–c) PCR and ELISA were used to detect Top 5 downregulated genes (TRIM34, DEFA3, DEFA1B, DEFA1, QPCT) and Top 5 upregulated genes (CHPT1, SMOX, FAM83A, GDF15, NAPG) in cord serum (a), maternal serum (b), and placental tissue (c). (d) The protein levels of Top 5 downregulated genes (TRIM34, DEFA3, DEFA1B, DEFA1, QPCT) and Top 5 upregulated genes (CHPT1, SMOX, FAM83A, GDF15, NAPG) were detected by western blot in placental tissues. ∗P<0:05 vs. control. N = 5 samples/group for the validation of DEGs.

Article Snippet: DEFA1 (CSB-E14155h, CUSABIO, China), QPCT (CSB-EL019135HU, CUSABIO, China), GDF15 (CSB-E12009h, CUSABIO, China), DEFA3 (CSB-EL006655HU, CUSABIO, China), DEFa1B (SEE136Hu, Cloud-Clone Corp, USA), and TRIM3 (SEC224Hu, CloudClone Corp, USA) kits were used for detection.

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Journal of Clinical Medicine

Article Title: Myocardial Involvement Detected Using Cardiac Magnetic Resonance Imaging in Patients with Systemic Sclerosis: A Prospective Observational Study

doi: 10.3390/jcm10225364

Figure Lengend Snippet: Correlations between changes in biochemical, CMR, and activity disease parameters.

Article Snippet: Growth/Differentiation Factor-15 (GDF-15; RayBiotech, Norcross, GA, USA) and galectin-3 (MyBiosource, San Diego, CA, USA) concentrations were determined using ELISA kits on a Nexgen ELISA four reader (Adaltis, Rome, Italy).

Techniques: Activity Assay