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R&D Systems quantikine elisa kit
Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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R&D Systems fibroblast growth factor 2
Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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R&D Systems recombinant human plateletderived growth factor
Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
Recombinant Human Platelet Derived Growth Factor, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Proteintech basic fibroblast growth factor
Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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Biosynth Carbosynth basic fibroblast growth factor
Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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R&D Systems fgf
Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by <t>ELISA</t> in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.
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Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by ELISA in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.

Journal: Journal of Bone Oncology

Article Title: Doxorubicin enhances adipogenesis in an FGF2-dependent manner and induces a tumour-promoting secretory phenotype

doi: 10.1016/j.jbo.2026.100754

Figure Lengend Snippet: Doxorubicin increases both intracellular and secreted protein levels of FGF2, and depletion of FGF2 impedes doxorubicin-enhanced adipogenesis. A) Protein expression of FGF2 at day 10 following treatment with vehicle control or 25 nM doxorubicin at day 9. Densitometry of FGF2 bands normalized to GAPDH as an endogenous loading control is shown for the high molecular weight (HMW) and low molecular weight (LMW) isoforms from 3 independent biological replicate blots. B) Secreted FGF2 (pg/mL) as detected by ELISA in conditioned supernatant at day 10 following vehicle control or 25 nM doxorubicin treatment at day 9 of adipogenic differentiation. C) Schematic for adipogenic differentiation of human MSC treated on day 9 of differentiation with vehicle control or 25 nM doxorubicin and subsequently transfected with 100 nM non-targeting siRNA or siRNA targeting FGF2 and assessed at day 12 by RT-qPCR and BODIPY staining. D) RNA isolated as per (C), was used to confirm siRNA-mediated depletion of FGF2 gene expression by RT-qPCR. E) Representative images of BODIPY stained cells treated as per (C) (left), with fold change in the percentage of BODIPY+ cells with doxorubicin treatment as compared to the vehicle control treatment for each siRNA condition enumerated and graphically represented (right). F) Gene expression of PPARG was measured using RT-qPCR at day 12 following treatment as in (C). Graphs show the mean ± SEM. Statistical tests are unpaired t -test in Fig A) and B) and one-way ANOVA in Fig D), E) and F) (*p < 0.05; **p < 0.01; ***p < 0.001), n = 3 biological replicates each with 3 technical replicates.

Article Snippet: ELISA to measure protein concentration of FGF2 in conditioned supernatants was performed using the Quantikine ELISA kit (cat #DFB50, R&D Systems, Minneapolis, MN), according to manufacturer’s instructions.

Techniques: Expressing, Control, High Molecular Weight, Molecular Weight, Enzyme-linked Immunosorbent Assay, Transfection, Quantitative RT-PCR, Staining, Isolation, Gene Expression