Journal: BMC Cancer

Article Title: Unveiling GDF15 as a promising biomarker for predicting survival in pancreatic ductal carcinoma: a retrospective research

doi: 10.1186/s12885-025-14963-7

Figure Lengend Snippet: IHC results of GDF15 level in PDAC and normal pancreas tissues. a Immunohistochemical analysis revealed markedly elevated GDF15 expression in pancreatic ductal adenocarcinoma (PDAC) specimens, whereas adjacent non‑neoplastic pancreatic tissues showed little to no detectable GDF15 staining. b Patients with poor prognoses exhibited higher expression levels compared to those with more favorable outcomes. c H‑score results for GDF15 expression were graphically represented, revealing a statistically significant difference in staining intensity between PDAC tumor tissues and their adjacent non‑neoplastic counterparts

Article Snippet: The TMAs were prepared using a PV-9000 IHC kit (ZSGB-BIO, Beijing, China) and incubated with an anti-GDF15 antibody (27455-1-AP; Proteintech, Wuhan, China).

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: BMC Cancer

Article Title: Unveiling GDF15 as a promising biomarker for predicting survival in pancreatic ductal carcinoma: a retrospective research

doi: 10.1186/s12885-025-14963-7

Figure Lengend Snippet: OS of PDAC patients with different GDF15 levels. Patients exhibiting low GDF15 expression showed significantly longer overall survival than those with high expression, as demonstrated by Kaplan–Meier analysis.

Article Snippet: The TMAs were prepared using a PV-9000 IHC kit (ZSGB-BIO, Beijing, China) and incubated with an anti-GDF15 antibody (27455-1-AP; Proteintech, Wuhan, China).

Techniques: Expressing

Journal: Molecular medicine (Cambridge, Mass.)

Article Title: CGREF1 modulates osteosarcoma proliferation by regulating the cell cycle through the Wnt/β-catenin signaling pathway.

doi: 10.1186/s10020-024-01038-9

Figure Lengend Snippet: Fig. 1 Expression level of CGREF1 in osteosarcoma was correlated with prognostic value for osteosarcoma patients. (A) The expression level of CGREF1 in various cancers was analyzed using data from the TIMER database. (B-C) Western blot assay was performed to detect the expression of CGREF1 in os teosarcoma tissues and adjacent normal tissues, and the results were summarized graphically based on three experiments. (D-E) Immunohistochemistry staining was conducted to examine the presence of CGREF1 in normal and osteosarcoma tissues, and a histogram format was used to summarize the results obtained from three experiments. (F) Kaplan-Meier survival analysis revealed that high expression levels of CGREF1 were associated with poor prognosis among osteosarcoma patients compared to those with low CGREF1 expression. (G-H) Western blot analysis was employed to determine the protein expression levels of CGREF1 in hFOB1.19, Saos-2, U2OS, 143B, and MG63 cell lines. Results from three experiments are summarized in a histogram format. Scale bar: 200 μm. (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: The Wnt pathway was pharmacologically inhibited using DIF-3 Keywords Osteosarcoma, CGREF1, Proliferation, Wnt/β-catenin signaling pathway, Cell cycle (MedChemExpress, USA) at a concentration of 30μM for subsequent experimental investigations. siRNA and plasmid transfection Specific small interfering RNA (siRNA) targeting CGREF1 and negative control siRNAs were purchased from OBiO Technology (Shanghai, China).

Techniques: Expressing, Western Blot, Immunohistochemistry, Staining

Journal: Molecular medicine (Cambridge, Mass.)

Article Title: CGREF1 modulates osteosarcoma proliferation by regulating the cell cycle through the Wnt/β-catenin signaling pathway.

doi: 10.1186/s10020-024-01038-9

Figure Lengend Snippet: Fig. 4 CGREF1 gene is implicated in the regulation of the Wnt signaling pathway. (A) Heatmap analysis was used to explore the expression pattern of CGREF1 related differentially expressed genes in RNA-seq data. (B) The KEGG enrichment analysis revealed a significant enrichment of CGREF1-associated genes in the Wnt signaling pathway. (C) The volcano plot depicted the differential gene expression and the expression relationship between cell cycle- related genes, wnt pathway-related genes and CGREF1 gene. (D) The GO enrichment analysis unveiled the involvement of CGREF1 in various cellular components, biological processes, and molecular functions

Article Snippet: The Wnt pathway was pharmacologically inhibited using DIF-3 Keywords Osteosarcoma, CGREF1, Proliferation, Wnt/β-catenin signaling pathway, Cell cycle (MedChemExpress, USA) at a concentration of 30μM for subsequent experimental investigations. siRNA and plasmid transfection Specific small interfering RNA (siRNA) targeting CGREF1 and negative control siRNAs were purchased from OBiO Technology (Shanghai, China).

Techniques: Expressing, RNA Sequencing, Gene Expression

Journal: Molecular medicine (Cambridge, Mass.)

Article Title: CGREF1 modulates osteosarcoma proliferation by regulating the cell cycle through the Wnt/β-catenin signaling pathway.

doi: 10.1186/s10020-024-01038-9

Figure Lengend Snippet: Fig. 7 CGREF1 knockdown inhibited the proliferation ability of osteosarcoma cells in vivo. (A) The tumor growth curve of nude mice subcutaneously injected with 143B-shCGREF1 and control cells. (B) The tumors of the nude mice in the two groups were isolated and photographed. (C) The tumors in the nude mice were weighed and compared. (D-E) Western blot was used to analyze the expression of CGREF1, β-catenin and Cyclin D in tumor tissues of the two groups. The results from three independent experiments were presented concisely in a histogram format. (F-G) IHC analysis of β catenin, Ki-67 and Cyclin D in xenograft tumors. The results from three independent experiments were presented concisely in a histogram format. Scale bar: 200 μm. (*P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: The Wnt pathway was pharmacologically inhibited using DIF-3 Keywords Osteosarcoma, CGREF1, Proliferation, Wnt/β-catenin signaling pathway, Cell cycle (MedChemExpress, USA) at a concentration of 30μM for subsequent experimental investigations. siRNA and plasmid transfection Specific small interfering RNA (siRNA) targeting CGREF1 and negative control siRNAs were purchased from OBiO Technology (Shanghai, China).

Techniques: Knockdown, In Vivo, Injection, Control, Isolation, Western Blot, Expressing

Journal: Molecular medicine (Cambridge, Mass.)

Article Title: CGREF1 modulates osteosarcoma proliferation by regulating the cell cycle through the Wnt/β-catenin signaling pathway.

doi: 10.1186/s10020-024-01038-9

Figure Lengend Snippet: Fig. 8 The regulatory mechanisms of CGREF1 on osteosarcoma proliferation

Article Snippet: The Wnt pathway was pharmacologically inhibited using DIF-3 Keywords Osteosarcoma, CGREF1, Proliferation, Wnt/β-catenin signaling pathway, Cell cycle (MedChemExpress, USA) at a concentration of 30μM for subsequent experimental investigations. siRNA and plasmid transfection Specific small interfering RNA (siRNA) targeting CGREF1 and negative control siRNAs were purchased from OBiO Technology (Shanghai, China).

Techniques:

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: Multifaceted validation of GDF15 changes in serum from SICM patients and their clinical associations. (A) GDF15 levels were quantified using the Luminex platform. (B) A volcano plot illustrated the gene expression distribution of GDF15 among differentially expressed genes (DEGs) in whole blood. (C) A heatmap displayed the expression profiles of GDF15 and inflammatory cytokines. (D) Serum GDF15 levels in patients. (E) Pearson correlation analysis demonstrated the association between GDF15 and SOFA score, as well as EF. (F) ROC curves were plotted to assess the diagnostic accuracy of GDF15 and SOFA score in identifying SICM. (G) Multivariate logistic regression analysis was performed to identify independent risk factors for the development of SICM in septic patients. ∗p < 0.05 indicates significant differences; ns: no significant differences.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Biomarker Discovery, Luminex, Gene Expression, Expressing, Diagnostic Assay

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: Upregulation of GDF15 in the SICM model. (A) A schematic workflow for the establishment of the SICM model in C57BL/6J mice via intraperitoneal injection of LPS or saline. (B) Cardiac contractile function parameters, including EF and FS. (C) Serum levels of GDF15 and IL-6. (D) Histopathological analysis of heart tissue, H&E staining (left) and immunohistochemical staining for Ly6G and CD68 (right). Black arrows indicate inflammatory cell infiltration; scale bar: 50 μm. (E) Western blot analysis of GDF15 protein expression in heart tissue. n = 4. (F) qPCR analysis of Gdf15 , Bnp , Il-1β , Il-6 , Icam-1 and Vcam- 1 mRNA levels in heart tissue. (G) Identification of GDF15-positive cells in single-cell RNA-sequencing dataset ( GSE190856 ). (H) qPCR analysis of Gdf15 and Il-1β , Il-6, Nos2, Ptgs2 mRNA expression in BMDM after LPS stimulation. ∗p < 0.05 indicates significant differences; n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Injection, Saline, Staining, Immunohistochemical staining, Western Blot, Expressing, RNA Sequencing

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: GDF15 deficiency exacerbates LPS-induced SICM in mice. (A) Schematic workflow for the establishment of the SICM model in Gdf15 −/− mice. Gdf15 −/− mice were intraperitoneally injected with LPS or saline to induce SICM, with tissue samples collected 24 h post-injection for further analysis. (B) Echocardiographic assessment of EF and FS. (C) H&E staining of heart tissue, black arrows indicate inflammatory cell infiltration. scale bar: 50 μm. (D) CD68 immunofluorescence staining of heart tissue. Blue staining highlights nuclei, red staining identifies CD68 + macrophages; scale bar: 20 μm. (E) qPCR analysis of mRNA expression levels of Bnp , Il-1β, Il-6, and Mcp-1 in heart tissue. n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Injection, Saline, Staining, Immunofluorescence, Expressing

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: MGP exerts anti-inflammatory effects via the MYPT1/AKT/YBX-1 signaling pathway. (A) IP-MS of BMDM to identify the interaction with GDF15. MYPT1 is marked in red. (B) Z-DOCK predicted the interaction domain between GDF15 and MYPT1. Pink represents GDF15, green represents MYPT1, and the boxed region indicates the binding domain. (C) Co-IP combined with Western blot analysis of GDF-15 and MYPT1 binding in macrophages after LPS treatment. (n = 3). (D) Immunofluorescence detection of co-localization between GDF15 (green) and MYPT1 (red), with blue staining for nuclei. Scale bar: 20 μm. (E) Protein expression levels of p -YBX-1, YBX-1, and p -AKT, AKT in BMDM after LPS and/or MGP treatment, with gray-scale intensity analysis of relative expression differences. (F) Representative immunofluorescence images of YBX-1 staining in BMDM after LPS and/or MGP treatment. Blue staining highlights nuclei, and red staining identifies YBX-1. Scale bar: 20 μm ∗p < 0.05, significantly different from control group. #p < 0.05, significantly different from LPS group. n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Protein-Protein interactions, Binding Assay, Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence, Staining, Expressing, Control

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: YBX-1 mediates GDF15-mediated transcriptional regulation of the NLRP3 pathway. (A) qPCR analysis of mRNA expression levels of Nlrp3, Asc , and Il-1β in LPS-stimulated BMDM after Si- Ybx-1 . (B) Western blot analysis of protein expression levels of NLRP3 and IL-1β in LPS-stimulated BMDM after YBX-1 knockdown. (C) qPCR analysis of mRNA expression levels of Nlrp3 and Il-1β in LPS and MGP-treated BMDM after YBX-1 knockdown. (D) Schematic diagram of the luciferase reporter plasmid for the Nlrp3 promoter. (E) Luciferase activity of pcDNA3.1-YBX-1 or empty vector-transfected cells. (F) Luciferase activity after LPS and MGP treatment. n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Expressing, Western Blot, Knockdown, Luciferase, Plasmid Preparation, Activity Assay, Transfection

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: Mechanism of action of macrophage-biomimetic nanocarriers delivering GDF15 to target the YBX-1-NLRP3 axis in SICM. Macrophage-biomimetic nanocarriers loaded with rhGDF15 are targeted to inflammatory sites in the heart, enhancing local drug accumulation, while GDF15 binds to MYPT1 to inhibit YBX-1 phosphorylation and block its nuclear translocation, leading to reduced nuclear YBX-1 expression and decreased transcriptional activity of the Nlrp3 promoter, which suppresses NLRP3 inflammasome assembly and pro-inflammatory cytokine release such as IL-1β, ultimately alleviating macrophage inflammatory responses, myocardial cell injury, and improving cardiac function in SICM.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Phospho-proteomics, Blocking Assay, Translocation Assay, Expressing, Activity Assay

Journal: Journal of Orthopaedic Surgery and Research

Article Title: URG4 mediates cell proliferation and cell cycle in osteosarcoma via GSK3β/β-catenin/cyclin D1 signaling pathway

doi: 10.1186/s13018-020-01681-y

Figure Lengend Snippet: Correlation analysis between URG4 protein expression and clinicopathological variables

Article Snippet: Slices were incubated with a URG4 primary antibody (1:100, Cat NO: 11998–1-AP, Proteintech, Wuhan, China) overnight at 4 °C, followed by further incubation with a secondary antibody (1:200, Cat NO: GB23303, Servicebio, Wuhan, China) at room temperature for 40 min.

Techniques: Expressing

Journal: Journal of Orthopaedic Surgery and Research

Article Title: URG4 mediates cell proliferation and cell cycle in osteosarcoma via GSK3β/β-catenin/cyclin D1 signaling pathway

doi: 10.1186/s13018-020-01681-y

Figure Lengend Snippet: Increased expression of URG4 in osteosarcoma tissues and cell lines. a URG4 expression was significantly increased in osteosarcoma tissues than corresponding normal tissues by HE and IHC, respectively (× 200 magnification). b PCR determined URG4 mRNA expression in osteosarcoma cell lines (HOS, MG63, Saos-2, U2OS, and 143B), and hFOB 1.19 was used as control. c Western blot assay determined URG4 protein expression in osteosarcoma cell lines (HOS, MG63, Saos-2, U2OS, and 143B), and hFOB 1.19 was used as control. d The mRNA expression level of the URG4 in HOS and MG63 cell lines following transfection as determined by RT-qPCR. e The protein expression level of URG4 in HOS and MG63 cell lines following transfection as determined by western blot assay. HE: hematoxylin and eosin; IHC: Immunohistochemistry; URG4: upregulated gene 4; Normal: normal tissues; OS: osteosarcoma tissues; K: blank group; NC: negative control. * p < 0.05, ** p < 0.01 vs the NC

Article Snippet: Slices were incubated with a URG4 primary antibody (1:100, Cat NO: 11998–1-AP, Proteintech, Wuhan, China) overnight at 4 °C, followed by further incubation with a secondary antibody (1:200, Cat NO: GB23303, Servicebio, Wuhan, China) at room temperature for 40 min.

Techniques: Expressing, Control, Western Blot, Transfection, Quantitative RT-PCR, Immunohistochemistry, Negative Control

Journal: Journal of Orthopaedic Surgery and Research

Article Title: URG4 mediates cell proliferation and cell cycle in osteosarcoma via GSK3β/β-catenin/cyclin D1 signaling pathway

doi: 10.1186/s13018-020-01681-y

Figure Lengend Snippet: URG4 affects vitro migration, invasion, colony formation, and vivo tumorigenesis. a HOS and MG63 cells stably transfected with siURG4 or siCtrl, and migration ability was evaluated by wound-healing assay. b Transwell assay was performed to analyze invasive ability. c Colony-forming abilities were researched by plate clones assay. d Images, volume, and weight of transplanted tumors in nude mice 31 days after subcutaneous inoculation with osteosarcoma cells stably expressing siURG4-1 or si-NC. URG4: upregulated gene 4; K: blank group; NC: negative control. * p < 0.05, ** p < 0.01 vs the NC

Article Snippet: Slices were incubated with a URG4 primary antibody (1:100, Cat NO: 11998–1-AP, Proteintech, Wuhan, China) overnight at 4 °C, followed by further incubation with a secondary antibody (1:200, Cat NO: GB23303, Servicebio, Wuhan, China) at room temperature for 40 min.

Techniques: Migration, Stable Transfection, Transfection, Wound Healing Assay, Transwell Assay, Clone Assay, Expressing, Negative Control

Journal: Journal of Orthopaedic Surgery and Research

Article Title: URG4 mediates cell proliferation and cell cycle in osteosarcoma via GSK3β/β-catenin/cyclin D1 signaling pathway

doi: 10.1186/s13018-020-01681-y

Figure Lengend Snippet: URG4 mediates proliferation, apoptosis, and cell cycle in vitro. a The proliferative capacity of cells following transfection was measured by CCK8 assay. b The apoptosis rate of HOS and MG63 cells was detected by flow cytometry. c The DNA content of PI-stained HOS and MG63 cells at the G0/G1, S, and G2/M phases was assessed by flow cytometry. CCK8: Cell Counting Kit-8; URG4: upregulated gene 4; NC: negative control. * p < 0.05, ** p < 0.01, *** p < 0.001 vs the NC

Article Snippet: Slices were incubated with a URG4 primary antibody (1:100, Cat NO: 11998–1-AP, Proteintech, Wuhan, China) overnight at 4 °C, followed by further incubation with a secondary antibody (1:200, Cat NO: GB23303, Servicebio, Wuhan, China) at room temperature for 40 min.

Techniques: In Vitro, Transfection, CCK-8 Assay, Flow Cytometry, Staining, Cell Counting, Negative Control

Journal: Journal of Orthopaedic Surgery and Research

Article Title: URG4 mediates cell proliferation and cell cycle in osteosarcoma via GSK3β/β-catenin/cyclin D1 signaling pathway

doi: 10.1186/s13018-020-01681-y

Figure Lengend Snippet: KEGG enrichment analysis of URG4-related gene

Article Snippet: Slices were incubated with a URG4 primary antibody (1:100, Cat NO: 11998–1-AP, Proteintech, Wuhan, China) overnight at 4 °C, followed by further incubation with a secondary antibody (1:200, Cat NO: GB23303, Servicebio, Wuhan, China) at room temperature for 40 min.

Techniques: Infection

Journal: Journal of Orthopaedic Surgery and Research

Article Title: URG4 mediates cell proliferation and cell cycle in osteosarcoma via GSK3β/β-catenin/cyclin D1 signaling pathway

doi: 10.1186/s13018-020-01681-y

Figure Lengend Snippet: URG4 regulates osteosarcoma cell growth by activating the GSK3β/β-catenin/cyclin D1 signaling pathway. a URG4-related genes enriched in the top 10 significant signaling pathway based on KEGG. b The protein level of GSK3β, p-GSK3β, nuclear β-catenin, and cyclin D1 in the HOS cell transfected with siURG4 was revealed by the western blot assay. c The protein level of GSK3β, p-GSK3β, nuclear β-catenin, and cyclin D1 in the MG63 cell transfected with siURG4 was revealed by the western blot assay. d The positive expression of the nuclear β-catenin protein in osteosarcoma cells was measured by the immunohistochemistry assay. e A model depicted URG4 promotes osteosarcoma progression via the GSK3β/β-catenin/cyclin D1 pathway. URG4: upregulated gene 4; NC: negative control. * p < 0.05 vs the NC

Article Snippet: Slices were incubated with a URG4 primary antibody (1:100, Cat NO: 11998–1-AP, Proteintech, Wuhan, China) overnight at 4 °C, followed by further incubation with a secondary antibody (1:200, Cat NO: GB23303, Servicebio, Wuhan, China) at room temperature for 40 min.

Techniques: Transfection, Western Blot, Expressing, Immunohistochemistry, Negative Control