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: Journal of Tissue Engineering

Article Title: GDF15 promotes osteogenic differentiation of human dental pulp stem cells by activating the TGF-β/SMAD signaling pathway

doi: 10.1177/20417314251357752

Figure Lengend Snippet: GDF15 secretion was significantly upregulated during the osteogenic differentiation of hDPSCs. (a) Morphology of primary hDPSCs. Scale bars (white): 100 μm. Morphology of passage 3 hDPSCs at 24 h of culture. Scale bars (black): 250 μm. P0 = Passage 0; P3 = Passage three. (b) Colony-forming assay to assess the self-renewal ability of hDPSCs. Scale bars: 250 μm. (c) The CCK-8 assay was employed to assess the proliferation of hDPSCs. (d) Proliferation of hDPSCs detected by crystal violet staining. Scale bars: 250 μm. (e) ALP staining of hDPSCs grown in OM for a week. Scale bars: 250 μm. (f) ARS staining of 21-day-cultured hDPSCs in OM and quantitative analysis of mineralized nodule deposition. Scale bars: 250 μm. (g) Flow cytometry demonstrated that the hDPSCs highly expressed CD105, CD90, and CD73; and lowly expressed CD45, CD19, and CD14. (h) During induction, intracellular mRNA levels of Gdf15 and osteogenic-specific genes ( Alp, Runx2, Osx, Ocn , and D spp ) were elevated. (i) During induction, osteogenic-specific proteins (ALP, RUNX2, OPN, DMP1, and DSPP) were upregulated and intracellular GDF15 protein was reduced in hDPSCs. Relative quantitative analysis of gray scale values of protein bands. The internal control was GAPDH. (j) Increased GDF15 secretion was detected by ELISA after osteogenic induction. Data were displayed as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.00001).

Article Snippet: The GDF15 antibody (Santa Cruz Biotechnology, Dallas, USA; sc-377195) was conjugated to protein A/G magnetic beads (MedChemExpress, New Jersey, USA) using a rotary mixer (Bio-Rad, Hercules, CA, USA) under sequential conditions: 30 min at room temperature and 2 h at 4°C.

Techniques: CCK-8 Assay, Staining, Cell Culture, Flow Cytometry, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Tissue Engineering

Article Title: GDF15 promotes osteogenic differentiation of human dental pulp stem cells by activating the TGF-β/SMAD signaling pathway

doi: 10.1177/20417314251357752

Figure Lengend Snippet: G df15 overexpression promoted osteogenic differentiation of hDPSCs in vitro , while Gdf15 knockdown suppressed it. (a) Images of GFP-positive hDPSCs were observed using fluorescence microscopy. Scale bars: 200 μm. GFP, Green Fluorescent Protein. (b) Relative mRNA expression of Gdf15 in the NC and G df15 overexpression ( Gdf15 ) groups. (c) Protein levels of G DF15 in the NC and Gdf15 groups. (d) Relative mRNA expression of G df15 in Gdf15 overexpressing hDPSCs after 7, 14, and 21 days of incubation in the OM. (e) ALP staining of the G df15 and NC groups on day 7 of osteogenic differentiation. (f) ARS staining and quantitative analysis of overexpressed hDPSCs were performed on days 7, 14, and 21 of osteogenic differentiation. (g) Osteogenic-specific genes ( Alp, Col1a1, Runx2, Osx , and Dmp1 ) and their relative mRNA levels in hDPSCs was examined by qPCR on days 7, 14, and 21 of the induced differentiation. (h) Relative mRNA expression of Gdf15 in the siNC and G df15 knockdown ( siGdf15 ) groups. (i) GDF15 protein levels in siNC and siGdf15 groups. (j) Relative mRNA expression of Gdf15 in siRNA-transfected hDPSCs after 7, 14, and 21 days of incubation in the OM. (k) ALP staining in the siNC and siGDF15 groups on day 7 of induced differentiation. (l) ARS staining and quantitative analysis were performed on hDPSCs transfected with siRNA on days 7, 14, and 21 of induced differentiation. (m) Relative mRNA levels of Alp, Col1a1, Runx2, Osx , and Dmp1 in siRNA-transfected hDPSCs on days 7, 14, and 21 of induced differentiation. Scale bars: 250 μm (ALP and ARS staining); Mean ± SD was employed to express all data (* p < 0.05, ** p < 0.01, *** p < 0.0001, **** p < 0.00001).

Article Snippet: The GDF15 antibody (Santa Cruz Biotechnology, Dallas, USA; sc-377195) was conjugated to protein A/G magnetic beads (MedChemExpress, New Jersey, USA) using a rotary mixer (Bio-Rad, Hercules, CA, USA) under sequential conditions: 30 min at room temperature and 2 h at 4°C.

Techniques: Over Expression, In Vitro, Knockdown, Fluorescence, Microscopy, Expressing, Incubation, Staining, Transfection

Journal: Journal of Tissue Engineering

Article Title: GDF15 promotes osteogenic differentiation of human dental pulp stem cells by activating the TGF-β/SMAD signaling pathway

doi: 10.1177/20417314251357752

Figure Lengend Snippet: rhGDF15 activated TGF-β/SMAD signaling pathway in hDPSCs. (a and b) Tgf-βr2 mRNA levels in hDPSCs overexpressing or knockdown of Gdf15 were measured after culturing in OM for 7, 14, and 21 days. (c) Tgf-βr2 mRNA levels in hDPSCs treated with OM containing rhGDF15 were measured after 7, 14, and 21 days of culture. (d) Structural modeling for molecular docking analysis of GDF15 and TGF-βR2. GDF15 and TGF-βR2 are depicted in blue-purple and orange-yellow, respectively. Hydrogen bonds are indicated in yellow. (e) Co-IP assay showing representative protein bands of GDF15 and TGFβ-R2 in hDPSCs. (f) Levels of TGF-β/SMAD signaling pathway-specific proteins after 20 ng/mL rhGDF15 treatment of hDPSCs for the indicated times were tested utilizing WB and grayscale analysis of protein bands. (g) Levels of TGF-β/SMAD signaling pathway-specific proteins after rhGDF15 (0–100 ng/mL) treatment of hDPSCs for 30 min were detected by WB and grayscale analysis of protein bands. (h) Expression of total, plasma, and nuclear proteins of p-SMAD2 and p-SMAD3 in hDPSCs after stimulation with rhGDF15 (20 ng/mL) for 30 min, and grayscale analysis of protein bands. (i) Immunofluorescence co-localization of TGF-β/SMAD signaling proteins in implants in the nude mouse subcutaneous transplantation model. Scale bars: 100 μm. The area of new bone formation is delineated by yellow dashed lines. The white dashed box demarcates the regions selected for high-magnification demonstration of osteogenic areas. Scale bars: 100 μm (enlarged view). Data were displayed as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The GDF15 antibody (Santa Cruz Biotechnology, Dallas, USA; sc-377195) was conjugated to protein A/G magnetic beads (MedChemExpress, New Jersey, USA) using a rotary mixer (Bio-Rad, Hercules, CA, USA) under sequential conditions: 30 min at room temperature and 2 h at 4°C.

Techniques: Knockdown, Co-Immunoprecipitation Assay, Expressing, Clinical Proteomics, Immunofluorescence, Transplantation Assay

Journal: Journal of Tissue Engineering

Article Title: GDF15 promotes osteogenic differentiation of human dental pulp stem cells by activating the TGF-β/SMAD signaling pathway

doi: 10.1177/20417314251357752

Figure Lengend Snippet: Activation of the TGF-β/SMAD signaling pathway by GDF15 is partially reversed by the inhibitors. (a) The concentration of the inhibitor was selected based on the analysis of the protein level ratios of phosphorylated to total SMAD2 or SMAD3. (b and c) The protein expression levels and quantitative analysis of p-SMAD2/SMAD2 and p-SMAD3/SMAD3 were assessed in hDPSCs pretreated with inhibitor for 1 h followed by rhGDF15 stimulation for 30 min. (d) Representative images and quantitative analysis of p-SMAD2 or p-SMAD3 immunofluorescence staining. Scale bars: 100 μm. (e and f) The osteogenic effects of inhibitor-treated hDPSCs were analyzed by qPCR. (g) ALP staining following 7 days of treatment. Scale bars: 250 μm. (h) ARS staining following 21 days of treatment and relative quantitative analysis. Scale bars: 250 μm. (i and j) The osteogenic effects of hDPSCs treated with inhibitors for 7 days were evaluated by WB, and the protein bands were analyzed in gray. Mean ± SD was employed to express all data (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: The GDF15 antibody (Santa Cruz Biotechnology, Dallas, USA; sc-377195) was conjugated to protein A/G magnetic beads (MedChemExpress, New Jersey, USA) using a rotary mixer (Bio-Rad, Hercules, CA, USA) under sequential conditions: 30 min at room temperature and 2 h at 4°C.

Techniques: Activation Assay, Concentration Assay, Expressing, Immunofluorescence, Staining