recombinant mouse fgf2 protein  (R&D Systems)

Journal: Blood

Article Title: Ferric carboxymaltose increases fracture risk in patients and reduces bone formation in mice with iron deficiency anemia

doi: 10.1182/blood.2025031806

Figure Lengend Snippet: FCM inhibits DMP1 binding to its potential cell surface receptor αVβ3-integrin and signaling. (A) Effect of 200 μg/mL of each IV iron formulation (FCM, FDI, or LMWID) on dose-dependent DMP1 binding to αVβ3-integrin. DMP1 bound to immobilized αVβ3-integrin was quantified immunometrically. Observed concentrations (y-axis) were plotted against increasing DMP1 concentrations (x-axis, ng/mL). (B) Binding of a constant DMP1 concentration (200 ng/mL) to immobilized αVβ3-integrin in the presence of increasing concentrations of FCM, FDI, or LMWID (10-200 μg/mL). (C) DMP1 binding to αVβ3 at pH 6.9, 7.4, and 7.9 in the additional presence of 200 μg/mL FCM or FDI. Binding is shown as a percentage of maximal αVβ3-DMP1 binding obtained at the highest DMP1 concentration in the absence of IV iron for each pH concentration. All data are shown as mean, n = 2. In panels A-C, a linear regression was applied, whereas in panel B, a nonlinear regression model was used. All data are shown as mean ± SD (n = 3). (D) Schematic illustration of the experimental approach used to assess recombinant murine DMP1-mediated signaling in MC3T3-E1 murine osteoblastic precursor cells by phosphoproteomics analysis. D created with BioRender.com . Wagner SA. (2026) https://BioRender.com/enm3jta . Cells were stimulated with 500 ng/mL DMP1 for 5 minutes, harvested in 100mM ammonium bicarbonate buffer, and processed for phosphoproteomics. (E) Phosphoproteomics data were analyzed with PANTHER Pathways. Enriched proteins were compared with the Mus musculus reference list, and the most significantly activated pathways are listed. Pathways further illustrated in panel F are marked in green (integrin signaling pathway) and blue (cytoskeletal regulation by Rho GTPase). (F) Illustration of the MAPK pathway and cytoskeletal regulation by Rho GTPase activated by DMP1 binding to αVβ3-integrin based on phosphoproteomics results. The MAPK signaling pathway and actin polymerization are initiated by the phosphorylation of focal adhesion kinase (FAK). F created with BioRender.com . Wagner SA. (2026) https://BioRender.com/enm3jta . (G) Activation of the MAPK pathway in MC3T3-E1 cells by DMP1 stimulation. Cells were incubated for 5 minutes with 500, 300, and 100 ng/mL of DMP1. FAK inhibitor (FAKi, 1 and 10μM) was used to block the phosphorylation cascade. Cell lysates were analyzed for phosphor-ERK1/2 (P-ERK) and total ERK 1/2 by western blot. α-Tubulin served as a loading control. (H) Representative western blot analysis of P-ERK, total ERK, and α-tubulin in MC3T3-E1 cell lysates after stimulation with 350 ng/mL DMP1 and 200 μg/mL IV iron formulations (FDI, FCM, LMWID, and IS) for 15 minutes. (I) Quantification of the western blot in panel H. (J) Western blot analysis of P-ERK, ERK, and α-tubulin in cell lysates after stimulation of MC3T3-E1 cells with 50 and 100 ng/mL FGF2 with or without 200 μg/mL IV iron formulations (FDI, FCM, and LMWID) for 15 minutes. (K) Representative western blot analysis of P-ERK, total ERK, and α-tubulin in U2OS cell lysates after stimulation with 500 ng/mL recombinantly expressed human C-terminal DMP1 and 200 μg/mL IV iron formulations (FDI, FCM, and LMWID) for 15 minutes. (L) RNA sequencing analysis of femora (bone marrow free) from mice aged 13 weeks, 1 week after 0.5 mg IV iron treatment, comparing Cntrl, IDA + NaCl, IDA + FCM, and IDA + FDI. Gene ontology analysis identified the integrin pathway as differentially regulated between groups. The heat map represents detected genes within this pathway, shown as z-scores and compared across all groups (mean of n = 3). Gene expression per mouse is shown in H. Arp2/3, actin related proteins 2 and 3; Cntrl, control; ERK1/2, extracellular signal-regulated kinase 1 and 2; Fe-NTA, iron-nitriloacetic acid; GRB2, growth factor receptor-bound protein 2; LC-MS/MS, liquid chromatography-tandem mass spectrometry; MEK, MAPK/ERK kinase; P-ERK, phosphorylated ERK; PI3K, phosphoinositide 3-kinase; PIP2, phosphatidylinositol 4,5-bisphosphate; Raf, rapidly accelerated fibrosarcoma–kinase; Ras-GTP, rat sarcoma bound to guanosine triphosphate; SD, standard deviation; SOS, son of sevenless homolog; TCA, tricarboxylic acid.

Article Snippet: Cells were stimulated for 15 minutes with 350 ng/mL murine DMP1 (R&D Systems, 4386-DM) or 50 ng/mL and 100 ng/mL FGF2 (Miltenyi Biotec; 130-105-787) with or without 200 μg/mL of FCM, FDI, or LMWID.

Techniques: Binding Assay, Cell Surface Receptor Assay, Formulation, Concentration Assay, Recombinant, Phospho-proteomics, Activation Assay, Incubation, Blocking Assay, Western Blot, Control, RNA Sequencing, Gene Expression, Liquid Chromatography with Mass Spectroscopy, Liquid Chromatography, Mass Spectrometry, Standard Deviation

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Schematic of the experimental setup with THP1 or Raw264.7 cells with indicated treatment. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p b Quantification of proportion of TREM2 in THP1 and Raw264.7 cells with PTX. n = 3 biological independent samples. c Western blot of THP1 and Raw264.7 cells with indicated treatments. The experiment was independently repeated three times with similar results. d Schematic of the experimental strategy. CM was collected from tumor cells with the indicated treatment. TREM2 expression in macrophages incubated with the CM was assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p e Quantification of proportion of TREM2 in THP1 incubated with CM of BT549 cells, SUM159 cells and MDA-MB-231 cells treated with PTX, in BMDM incubated with CM of Py8119 treated with PTX, in Raw264.7 incubated with CM from 4T1 and Py8119 cells treated with PTX, respectively. n = 3 biological independent samples. f Western blot analysis of TREM2 and related proteins in THP1, BMDMs, and Raw264.7 cells incubated with CM from BT549, SUM159, Py8119, and 4T1 cells, respectively. The experiment was independently repeated three times with similar results. g Representative immunofluorescence staining of THP1 cells incubated with BT549 CM. n = 3 biological independent samples. h Quantification of proportion of TREM2 in THP1 incubated with CM from BT549, SUM159, and MDA-MB-231 cells treated with Nab-PTX, respectively. n = 3 biological independent samples. i Quantification of the proportion of TREM2 in BMDM incubated with CM of Py8119 treated with Nab-PTX. n = 3 biological independent samples. j Cytokine array analysis of CM from BT549 cells treated indicated treatment. k Western blot analysis of the indicated proteins in Raw264.7 cells and BMDMs treated with recombinant FGF2. The experiment was independently repeated three times with similar results. l Quantification of the proportion of TREM2 in Raw264.7 cells and BMDMs treated with recombinant FGF2. n = 3 biological independent samples. m Schematic of the experimental strategy. CM was collected from tumor cells with the indicated treatment, and then pretreatment using an FGF2 neutralizing antibody. TREM2 expression in macrophages incubated with the CM was assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p n Western blot analysis of the indicated proteins in Raw264.7 and BMDMs incubated with the indicated CM. The experiment was independently repeated three times with similar results. o Quantification of the proportion of TREM2 in Raw264.7 and BMDMs incubated with the indicated CM. n = 3 biological independent samples. p Representative multiplex immunofluorescence staining of CD68, TREM2 and FGF2 in tumors treated with PTX or Nab-PTX ( n = 3 mice per group). The experiment was independently repeated three times with similar results. Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( b , e , h and i ) and two-sided one-way ANOVA followed by Tukey’s test ( l and o ). Source data are provided as a Source Data file.

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques: Western Blot, Expressing, Incubation, Immunofluorescence, Staining, Recombinant, Multiplex Assay

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Overlap of RNA-seq ( n = 3) and PROMO public database analyses to predict transcription factors regulating TREM2 expression. b Correlations between FGF2 and ATF3 expression in breast cancer using TCGA databases. c qPCR analysis of Atf3 expression in the indicated cells. n = 4 (DMSO and PTX) or 3 (PBS and Nab-PTX) biological independent samples. d Western blot analysis of the indicated proteins in Py8119 cells transfected with ATF3-expressing or control vectors. The experiment was independently repeated three times with similar results. e qPCR analysis of Fgf2 expression in Py8119 cells transfected with Atf3 -expressing or control vectors. n = 3 biological independent samples. f Luciferase activity in HEK293T cells transfected with the indicated reporters and Atf3 -expressing or control vectors. n = 3 biological independent samples. g Abundance of Atf3 bound to the Trem2 promoter in Py8119 cells, as assessed by ChIP-qPCR. n = 3 biological independent samples. h ATAC-seq tracks showing the chromatin accessibility in the ATF3 loci for BT549 cells treated by PTX or Nab-PTX. n = 2 samples per group. Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( c, e and f ) and two-sided two-way ANOVA followed by Šídák’s test ( g ). Source data are provided as a Source Data file.

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques: RNA Sequencing, Expressing, Western Blot, Transfection, Control, Luciferase, Activity Assay, ChIP-qPCR

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: a Overlap of GeneCards and EDCODE public databases analyses to predict transcription factors that regulate TREM2. b Western blot analysis of the indicated proteins in BMDMs and Raw264.7 cells incubated with FGF2 (left) and with CM from Py8119 cells treated with PTX (right). The experiment was independently repeated three times with similar results. c Western blot analysis of the indicated proteins in Raw264.7 cells incubated with the indicated treatment. The experiment was independently repeated three times with similar results. d qPCR analysis of Trem2 expression in BMDMs transfected with Egr1 -expressing vectors. n = 3 biological independent samples. e Western blot analysis of TREM2 expression in BMDMs transfected with Egr1 -expressing vectors. The experiment was independently repeated three times with similar results. f Luciferase activity of HEK293T cells transfected with the indicated reporters and EGR1 -expressing or control vectors. n = 3 biological independent samples. g Abundance of EGR1 bound to the TREM2 promoter in BMDMs assessed by ChIP-qPCR. n = 3 biological independent samples. h Schematic of the Transwell assay. The first CM was collected from tumor cells with indicated treatment, and the second CM was collected from macrophages incubated with the first CM. The migration and invasion capabilities of macrophages incubated with the first CM were assessed. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p i Quantification of migration and invasion of Py8119 cells induced by BMDM CM incubated with CM from Py8119 cells treated with PTX, and BT549, SUM159, and MDA-MB-231 cells induced by THP1 CM incubated with CM from BT549, SUM159, and MDA-MB-231 cells treated with PTX (left) or Nab-PTX (right), respectively. n = 5 biological independent samples. j Cytokine array analysis of CM from BMDMs with or without TREM2. k Quantification of migration and invasion of Py8119 cells incubated with indicated proteins. n = 3 biological independent samples. l Quantification of migration and invasion of Py8119 cells induced by CM from BMDMs ( Trem2 +/+ ) with CDE-096. n = 3 biological independent samples. m Western blot analysis of EMT- stimulating proteins in BMDMs incubated with indicated proteins. The experiment was independently repeated three times with similar results. n Schematic illustration showing that FGF2 promotes ERK1/2 phosphorylation to upregulate EGR1, which increases TREM2 expression in macrophages. Upregulated TREM2 enhances the secretion of Serpin E1, HGF, CCL3, and CXCL2 from macrophages to tumor cells, facilitating tumor metastasis via EMT. Created in BioRender. Xing, Y. (2026) https://BioRender.com/nsp747p . Data are shown as means ± S.D. and were analyzed by two-sided unpaired Student’s t test ( d, f, i and l ), two-sided one-way ANOVA followed by Tukey’s test ( k ) and two-sided two-way ANOVA followed by Šídák’s test ( g ). Source data are provided as a Source Data file.

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques: Western Blot, Incubation, Expressing, Transfection, Luciferase, Activity Assay, Control, ChIP-qPCR, Transwell Assay, Migration, Phospho-proteomics

Journal: Nature Communications

Article Title: Paclitaxel drives TREM2 + macrophage expansion underlying its inferior therapeutic efficacy compared to Nab-paclitaxel

doi: 10.1038/s41467-026-69060-5

Figure Lengend Snippet: PTX, but not Nab-PTX, promotes lung metastasis by inducing TREM2 + macrophage recruitment. Mechanistically, PTX enhances the ATF3-FGF2 axis in breast cancer cells; secreted FGF2 activates the EGR1–TREM2–EMT cytokine axis in macrophages. Created in BioRender. Xing, Y. (2026) https://BioRender.com/6hxlbow .

Article Snippet: The FGF2 protein (HY-P73052AF) was purchased from MedChemExpress and used with indicated concentration.

Techniques:

Journal: Stem Cells Translational Medicine

Article Title: FGF2 alters the calvarial suture niche homeostasis shifting skeletal stem cell/progenitors toward an osteo-angiogenic coupling fate

doi: 10.1093/stcltm/szag003

Figure Lengend Snippet: Morphology of FGF-treated calvarial suture explants. A, Schematic outlining the experimental procedure. B, Phase contrast images of a representative calvarial suture (COR suture) explant after 8 days in culture with recombinant FGF2 protein (100 ng/mL) revealing the presence of a tube-like structure (red arrows) sprouting from the edge (green arrow) of the suture (magnification at 10×). C, 20× magnification of the red dashed boxed area in panel B. Panel D highlights the presence of round cells; bottom panel D(i) is a magnification at 40× of the boxed area in panel D, and green arrows mark the small, round, and bright cells. E, Phase contrast image of FGF2 treated PF suture shows a morphology similar to FGF2 treated COR suture. Green arrows point to the suture edge. Red arrows mark tube-like structures and blue arrow a lacunae-like formation. COR calvarial suture explant control. Green arrows point the suture edge. Magnification at 20×. F, Phase contrast image of untreated COR calvarial suture explant control. G, Phase contrast image of untreated PF calvarial suture explant control. Green arrows point the suture edge. H, Cartoon depicting the following experimental steps. I-K, Representative phase contrast images of cells isolated by FACS sorting from FGF2 treated COR suture explants cultured in presence of FGF2 (100 ng/mL), followed by VEGF (150 ng/mL) addition at day 4 for additional 48 hours (total 6 days of culture). I, Cells display a foci formation (dashed green circle) and sprouting of tubular structures in panels J and K, (green arrows). Magnification at 20× and 40×. L, Control cells isolated from untreated calvarial suture explants grow in a monolayer without forming structures resembling either foci or tube-like structures (magnification at 20×). Experiments were performed at least 3 times. Scale bars: 100 µm in panel B; 50 µm panels (C, D, E, F, G, I, J, I, L); 20 µm in panel D(i) and K. Abbreviations: COR, coronal; FGF, fibroblast growth factor.

Article Snippet: Recombinant mouse FGF2 protein (#3139, R&D System) was added at concentration of 100 ng/mL.

Techniques: Recombinant, Control, Isolation, Cell Culture

Journal: Stem Cells Translational Medicine

Article Title: FGF2 alters the calvarial suture niche homeostasis shifting skeletal stem cell/progenitors toward an osteo-angiogenic coupling fate

doi: 10.1093/stcltm/szag003

Figure Lengend Snippet: Bulk RNA-Seq profiling of skeletal stem cell/progenitor population isolated from FGF2 treated and untreated sutures. A and B, Heatmaps of endothelial cell and endothelial progenitor cell marker genes, highlight a distinct upregulation of endothelial and endothelial progenitor cells (EPC) in FGF2 treated cells. Red color: upregulation (≥3-fold); Green color: downregulation. (−≥3-fold). The (+) symbol points to genes that are upregulated in the cell populations analyzed. C, Heatmap of osteogenic marker downregulation in cells as above. D, Validation of up- and down-regulation of key endothelial and osteogenic markers by RT-PCR analysis. E, Diagram/cartoon summarizing the trend in the expression profiles of key endothelial and osteogenic markers up- or downregulated by (≥3-fold) in cells isolated from FGF2 treat and untreated sutures.

Article Snippet: Recombinant mouse FGF2 protein (#3139, R&D System) was added at concentration of 100 ng/mL.

Techniques: RNA Sequencing, Isolation, Marker, Biomarker Discovery, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Stem Cells Translational Medicine

Article Title: FGF2 alters the calvarial suture niche homeostasis shifting skeletal stem cell/progenitors toward an osteo-angiogenic coupling fate

doi: 10.1093/stcltm/szag003

Figure Lengend Snippet: Chromatin accessibility between skeletal stem cells/progenitors derived from FGF2 treated and untreated calvarial sutures. A, Representation of enriched chromatin active motifs identified in skeletal stem cell/progenitor population isolated from FGF2 treated calvarial suture explants and predicted to function as activated transcription sites. Associated transcription factors and target genes are listed. B, Heatmap of peaks differentially expressed in cell isolated from FGF2 treated suture explants and untreated suture explants. C, Pie charts showing the proportion of chromatin open regions. D, Go-enrichment of biological processes in cells isolated from FGF2 treated suture explants unveiling blood vessel morphogenesis as major enriched process. E, KEGG enrichment analysis also reveals processes associated to angiogenesis, such as hemostasis, developmental biology, and extracellular matrix remodeling. F, Reactome enrichment analysis showing the gene ratio/intensity related to the biological processes listed in E.

Article Snippet: Recombinant mouse FGF2 protein (#3139, R&D System) was added at concentration of 100 ng/mL.

Techniques: Derivative Assay, Isolation

Journal: Stem Cells Translational Medicine

Article Title: FGF2 alters the calvarial suture niche homeostasis shifting skeletal stem cell/progenitors toward an osteo-angiogenic coupling fate

doi: 10.1093/stcltm/szag003

Figure Lengend Snippet: CITE sequencing analysis of skeletal stem cell/progenitor population isolated from FGF2 treated and untreated calvarial sutures. A, UMAP analysis showing changes in the representation between cells isolated from untreated (control) and FGF2 treated calvarial suture explants. B, Pies illustrating cluster-ratio overview of cell subpopulations isolated from untreated (control) and FGF2 treated suture explants (yellow lines mark clusters increased in cell representation; purple lines clusters with decreased cell representation) C, CITE-Seq Heatmap of the surface markers CD31 (PECAM-1), CD133 (Prominin-1), Sca-1 and CD200 reveals their increased levels in cells derived from FGF2 treated sutures (yellow indicates downregulation; purple upregulation). D, Violin plots showing the distribution of cells expressing the four CITE-Seq markers (top panel) and histogram of gene module scores expression (bottom panel) in control and FGF2 treated populations. E, Violin plots showing the distribution of cells expressing the four CITE-Seq markers (upper panel) in all 10 clusters and histogram of genes module scores (bottom panel). F, CITE-Seq heatmap of the surface markers CD31 (PECAM-1), CD133 (Prominin-1), Sca-1 and CD200 in all 10 clusters. G, Illustration of RNA clusters of cells expressing key markers of the osteogenic lineage by Violin plots (left panel), osteogenic modules (middle panel), and heatmap (right panel). H, Illustration of RNA clusters of cells expressing key markers of the endothelial/angiogenic lineage by Violin plots (left panels), angiogenic modules (top right panel), and heatmap (bottom right panel). I, Cartoon illustrates inferred trajectories/directions of lineages differentiation. J, Shows the velo latent time of differentiation for the different clusters. Yellow color indicates the more differentiated status of the cell populations.

Article Snippet: Recombinant mouse FGF2 protein (#3139, R&D System) was added at concentration of 100 ng/mL.

Techniques: Sequencing, Isolation, Control, Derivative Assay, Expressing

Journal: Stem Cells Translational Medicine

Article Title: FGF2 alters the calvarial suture niche homeostasis shifting skeletal stem cell/progenitors toward an osteo-angiogenic coupling fate

doi: 10.1093/stcltm/szag003

Figure Lengend Snippet: Skeletal stem cell/progenitor population isolated from FGF2 treated suture explants display an endothelial functional profile. A, Confocal microphotographs of an in vitro Matrigel assay performed on a pool of cells isolated from FGF2 treated PF and COR sutures and untreated sutures. Tube-like structures are observed after 18 hours. Staining dye (green color) shows the endothelial tube formation (left panel). Middle panel showing absence of tube-like structure formation in cells isolated from untreated calvarial sutures explants. Addition of the inhibitor vinblastine suppresses tube-like formation (right panel) in cells isolated from FGF2 treated suture explants. Experiments were performed three times. B, Quantification of in vitro tube-like formation by length measurement using ImageJ2 program. Results are presented as mean ± SD of tube-length formation. Analysis was performed on four independent assays. A significant difference in tube-formation length is observed in FGF2 treated cells. C, Acetylated LDL uptake by pool of cells isolated from FGF2 treated PF and COR sutures is confirmed by staining of DiI dye (left panel), whereas cells isolated from untreated calvarial suture explants do not uptake Acetylated LDL (middle panel). Likewise, in the presence of the inhibitor Protein-S (P-S), cells isolated from FGF2 treated sutures do not uptake acetylated LDL (right panel). D, Quantification of acetylated LDL uptake using ImageJ2 program (ImageJ 2, NIH) confirms a significant increased uptake by FGF2 treated cells as compared to untreated and P-S treated cells. Densitometric results are mean ± SD of the threshold levels from two independent experiments. E, H&E staining of OCT cryo-tissue sections from in vivo Matrigel plug assay showing the ability of cells isolated from FGF2 treated sutures to form vessels (left panel) which stained positive for CD31/PECAM-1 (green color) as assessed by immunofluorescence using specific antibodies (middle panel). Immunostaining using Sca-1 antibodies (red color), a marker associated with endothelial progenitor cells (EPC), reveals the presence of positive cells (right panel), and their partial colocalization with CD31/PECAM-1 positive cells as shown in panel F. G, Matrigel plug assay performed with cells isolated from untreated calvarial sutures does not reveal any vessel formation. Scale bars: 100 µm in panels A and C; 50 µm in panels E and G; 20 µm in panel F.

Article Snippet: Recombinant mouse FGF2 protein (#3139, R&D System) was added at concentration of 100 ng/mL.

Techniques: Isolation, Functional Assay, In Vitro, Matrigel Assay, Staining, In Vivo, Immunofluorescence, Immunostaining, Marker

Journal: Stem Cells Translational Medicine

Article Title: FGF2 alters the calvarial suture niche homeostasis shifting skeletal stem cell/progenitors toward an osteo-angiogenic coupling fate

doi: 10.1093/stcltm/szag003

Figure Lengend Snippet: Osteo-induction assay of skeletal stem cell/progenitor population isolated from FGF2 treated and untreated suture explants. A and B, Contrast-phase microphotographs of an osteogenic assay stained with von-Kossa procedure after 21 days in culture with osteogenic medium. Cells isolated from FGF2 treated suture explants (pool of PF and COR sutures) form a very dense and patchy mineralized areas as revealed by von Kossa staining (magnification at 20×). Right panels are a magnification at 40× of the red dashed boxed areas in panel A and B, green arrows mark the presence of tube-like structures. C, Von Kossa staining of cells derived from untreated suture explants showing a diffuse ECM mineralization and some bone nodules (yellow arrows) (magnification at 20×). Right panel is a magnification at 40× of the red dashed boxed area in panel C. D, RT-PCR analysis of osteogenic and endothelial markers profiles expression of both osteogenic and angiogenic markers in cells isolated from FGF2 treated suture explants (top panel), cells express all four markers. E, Cells isolated from untreated suture explants (pool of PF and COR sutures) express less of the osteogenic marker Runx2 and low levels of Bglap and Ve-cadherin, while no expression of the endothelial marker CD3 1 ( Pecam1 ) is detected (bottom panel). Experiments were performed two times. Scale bars: 100 µm in panels A-C (left side); 50 µm in panels A-C (right side). F, Densitometric analysis of the PCR bands performed by ImageJ2 program (ImageJ 2, NIH). The densitometric results were normalized to their respective loading control ( Gapdh -bands) and presented as percent increase (100%). Results are mean ± SD of three independent analyses. Statistical significance: * P <.05.

Article Snippet: Recombinant mouse FGF2 protein (#3139, R&D System) was added at concentration of 100 ng/mL.

Techniques: Isolation, Staining, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Marker, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Development of a biomimetic thyroid acellular scaffold as a 3D platform for modeling thyroid cancer aggressiveness and drug resistance

doi: 10.3389/fbioe.2025.1692549

Figure Lengend Snippet: Retention of endogenous growth factors in the Thyroid Acellular Scaffold (TAS). The bar chart shows the concentrations (pg/mL) of key growth factors—Vascular Endothelial Growth Factor (VEGF), Transforming Growth Factor-β (TGF-β), Hepatocyte Growth Factor (HGF), Fibroblast Growth Factor (FGF), Epidermal Growth Factor (EGF), and Platelet-Derived Growth Factor (PDGF)—in native thyroid tissue and the TAS, as quantified by ELISA. Although the decellularization process resulted in a significant reduction in the concentration of all measured growth factors, the TAS successfully retained a substantial amount of these bioactive molecules. Data are presented as mean ± standard deviation. **p < 0.01 compared to native tissue.

Article Snippet: Specific ELISA kits were used to detect the content of vascular endothelial growth factor (VEGF, R&D Systems, RRV00), transforming growth factor-β (TGF-β, R&D Systems, RTB100B), hepatocyte growth factor (HGF, R&D Systems, MHG00), fibroblast growth factor (FGF, R&D Systems, MFB00), epidermal growth factor (EGF, R&D Systems, DY3214), and platelet-derived growth factor (PDGF, R&D Systems, MBB00).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Standard Deviation