Journal: Journal of Clinical Medicine

Article Title: The Influence of Hypertensive Therapies on Circulating Factors: Clinical Implications for SCFAs, FGF21, TNFSF14 and TNF-α

doi: 10.3390/jcm9092764

Figure Lengend Snippet: ( a ) Antihypertension treatments decreased the level of FGF21 in hypertensive patients ( n = 12–18). ( b ) Circulating levels of FGF21 increase with age ( n = 5–14) and ( c ) BMI ( n = 3–21). All data are presented as the mean ± SEM. FGF21, fibroblast growth factor 21.

Article Snippet: Serum was analysed for FGF21, TNFSF14, TNF-α and insulin, using the appropriate enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s respective instructions (Duoset human FGF21 ELISA kit (Cat# DY2539, R&D systems, Inc., Minneapolis, MN, USA), Duoset human TNFSF14 ELISA kit (Cat# DY664, R&D systems) and Duoset human TNF-α ELISA kit (Cat# DY210, R&D systems)).

Techniques:

Journal: Endocrine Connections

Article Title: Circulating FGF21 is lower in South Asians compared to Europids with type 2 diabetes mellitus

doi: 10.1530/ec-24-0362

Figure Lengend Snippet: Figure 2. Comparison of serum fibroblast growth factor 21 concentration in South Asian and Europid males and females with type 2 diabetes mellitus. Box plots showing serum concentration of fibroblast growth factor 21 (FGF21) in all South Asians (n=47, orange circles) compared to all Europids (n=49, green circles) with type 2 diabetes mellitus (A). In addition, FGF21 concentrations are shown for South Asian (n=19, orange circles) compared to Europid (n=29, green circles) males (B) and South Asian (n=28, orange circles) compared to Europid (n=20, green circles) females (C). Circles represent individual values, boxes represent means, and deviations represent standard deviations.

Article Snippet: Serum fibroblast growth factor 21 (FGF21) concentrations in samples from all three trials were measured using the human FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA).

Techniques: Comparison, Concentration Assay

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Concentration-dependence of BrdU incorporation in A2B5+ OPCs cultured with adult mice serum (n = 6). (B) BrdU incorporation in A2B5+ OPCs 1 day after stimulation with adult mouse serum heated or pretreated with the indicated reagents (n = 6). (C) BrdU incorporation in OPCs after serum stimulation with PD173074 (10 nM), an inhibitor of FGFR (n = 4). (D) BrdU incorporation in OPCs after serum stimulation with NF449 (10 μM), an inhibitor of FGFR3 (n = 4). (E) BrdU incorporation in mouse OPCs with FGFR and β-klotho knockdown after serum stimulation (n = 7). (F) BrdU incorporation in OPCs after stimulation with recombinant FGF15, FGF21, and FGF23 (n = 4). (G) BrdU incorporation in OPCs after serum stimulation with neutralizing antibody against FGF21 (n = 5), determined by Student’s t test or by ANOVA with Tukey’s post hoc test or Dunnett’s test. Error bars represent SEM. *P < 0.05, **P < 0.01.

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Concentration Assay, BrdU Incorporation Assay, Cell Culture, Recombinant

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Quantitations of Fgf21 mRNA (left) and FGF21 protein (right) in individual organs of intact mice (n = 9); **P < 0.01. (B) Representative images of FGF21-immunolabeled pancreas of intact mice (n = 3). (C) Double IHC staining for FGF21 with the indicated cell markers in the pancreas of adult mice (n = 3). (D) BrdU incorporation in mouse OPCs after stimulation with serum from mice with FGF21 knockdown in the pancreas (n = 4); *P < 0.05, **P < 0.01, as determined by ANOVA with Tukey’s post hoc test. Error bars represent SEM. Scale bars: 50 μm (B); 10 μm (C).

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Immunolabeling, Immunohistochemistry, BrdU Incorporation Assay

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Quantitation of FGF21 protein in the spinal cord 1 day and 3 days after LPC injection (n = 5 for control, 5 for d1, 4 for d3). (B) Representative images of spinal cord sections double-labeled for PDGFRα and Ki67. Sections were obtained from FGF21-KO mice and control littermates 7 days after LPC injection. Graph shows quantitations as indicated in the images (n = 5). Arrows indicate Ki67+ cells colabeled with PDGFRα; arrowheads indicate Ki67+ cells not labeled with PDGFRα. (C) Representative images of spinal cord sections labeled for MBP. Sections were obtained from mouse spinal cord 14 days after LPC injection. Graph shows quantitations as indicated in the images (n = 5). (D) Representative immunoelectron microscopy images of myelin in the spinal cord. Sections were obtained from FGF21-KO mice and control littermates 14 days after LPC injection. Graphs show quantitations of g-ratio indicated in the images (n = 3). (E) Motor function was assessed by ladder-walk test (n = 11 for control littermates, 9 for FGF21-KO mice). (F) Representative images of spinal cord sections labeled for MBP. Graph shows quantitations as indicated in the images (n = 5 for control littermates + vehicle, 5 for FGF21-KO mice + vehicle, 4 for FGF21-KO mice + FGF21). (G) Representative images of spinal cord sections labeled for MBP. Graph shows quantitations as indicated in the images (n = 4); **P < 0.01 as determined by Student’s t test or by ANOVA with Tukey’s post hoc test or Dunnett’s test. *P < 0.05, **P < 0.01. Error bars represent SEM. Scale bars: 50 μm (B); 200 μm (C, F, and G); 2 μm (D).

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Quantitation Assay, Injection, Labeling, Immuno-Electron Microscopy

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Representative images of β-klotho expression in the mouse spinal cord 3 days after LPC injection. (B) Representative images of spinal cord sections double-labeled for PDGFRα and Ki67. Graph shows quantitations as indicated in images (n = 3 for control, 3 for CKO); *P < 0.05. Arrows indicate Ki67+ cells colabeled with PDGFRα. (C) Representative images of spinal cord sections labeled for MBP. Graph shows quantitations as indicated in the images (n = 4 each); **P < 0.01. (D) Representative images of brain sections double-labeled for PDGFRα and Ki67 in the mouse cortex, 7 days after traumatic brain injury. FGF21 was administered i.c.v. 24 hours after LPC injection (n = 4); *P < 0.05. Arrows indicate Ki67+ cells colabeled with PDGFRα; arrowheads indicate Ki67+ cells not labeled with PDGFRα. (E) Representative images of brain sections labeled for MBP in the mouse cortex, 14 days after traumatic brain injury. Graph shows quantitations as indicated in the images (n = 6 for vehicle, n = 4 for FGF21); *P < 0.05. (F) Representative immunoelectron microscopy images of myelin in the mouse cortex, 14 days after traumatic brain injury. Graphs show quantitations of g-ratio indicated in the images (n = 4); **P < 0.01 as determined by Student’s t test. Error bars represent SEM. Scale bars: 20 μm (A); 50 μm (B and D); 200 μm (C and E); 2 μm (F).

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Expressing, Injection, Labeling, Immuno-Electron Microscopy

Journal: The Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/JCI94337

Figure Lengend Snippet: (A) Representative image of β-klotho expression in an autopsied sample from healthy patient and a patient with multiple sclerosis. Graphs show quantitations as indicated in the images (n = 4 for healthy patients, 3 for multiple sclerosis patients); **P < 0.01. (B) BrdU incorporation in human OPCs after stimulation with recombinant FGF21 (n = 6 for control, 4 for FGF21); *P < 0.05 as determined by Student’s t test. Error bars represent SEM. Scale bar: 20 μm.

Article Snippet: To assess cell proliferation, cells were cultured in DMEM supplemented with or without recombinant human FGF21 (R&D Systems) at a final concentration of 6 μg/ml.

Techniques: Expressing, BrdU Incorporation Assay, Recombinant

Journal: Biochemical Journal

Article Title: Heparan sulfate 6-O-endosulfatases: discrete in vivo activities and functional co-operativity

doi: 10.1042/bj20060848

Figure Lengend Snippet: Figure 6 Loss of mSulf activity increases the mitogenic response of MEFs to FGF2

Article Snippet: Recombinant human FGF2 and HGF/SF (HGF/scatter factor) were from R&D Systems (Abingdon, Oxon., U.K.).

Techniques: Activity Assay

Journal: Molecules

Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans

doi: 10.3390/molecules24183360

Figure Lengend Snippet: Figure 3. Bar graphs of normalized different growth factors binding preference to surface heparin by competing with different size of heparin oligosaccharides (from fp4 to dp18). (A) FGF2, (B) FGF7, (C) FGF10, (D) HGF, (E) TGFβ-1. Concentrations were 200, 100, 100, 20, and 50 nM for FGF2, FGF7, FGF10, HGF and TGFβ-1, respectively, and concentrations of heparin oligosaccharides were 1000 nM. All bar graphs based on triplicate experiments.

Article Snippet: Recombinant human FGF2 was a gift from Amegen; FGF7 and FGF10 were generously provided by Professor Mohammadi from NYU; HGF and TGF β-1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: Molecules

Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans

doi: 10.3390/molecules24183360

Figure Lengend Snippet: Figure 4. Bar graphs of normalized different growth factors binding preference to surface heparin by competing with different chemical modified heparin in solution. (A) FGF2, (B) FGF7, (C) FGF10, (D) HGF, (E) TGFβ-1. Concentrations were 200, 100, 100, 20, and 50 nM for FGF2, FGF7, FGF10, HGF and TGFβ-1, respectively, and concentrations of different modified heparin were 1000 nM. All bar graphs based on triplicate experiments.

Article Snippet: Recombinant human FGF2 was a gift from Amegen; FGF7 and FGF10 were generously provided by Professor Mohammadi from NYU; HGF and TGF β-1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: Molecules

Article Title: Comparison of the Interactions of Different Growth Factors and Glycosaminoglycans

doi: 10.3390/molecules24183360

Figure Lengend Snippet: Figure 5. Bar graphs of normalized different growth factors binding preference to surface heparin by competing with different GAGs in solution. (A) FGF2, (B) FGF7, (C) FGF10, (D) HGF, (E) TGFβ-1. Concentrations were 200, 100, 100, 20, and 50 nM for FGF2, FGF7, FGF10, HGF and TGFβ-1, respectively and concentrations of GAGs were 1000 nM. All bar graphs based on triplicate experiments.

Article Snippet: Recombinant human FGF2 was a gift from Amegen; FGF7 and FGF10 were generously provided by Professor Mohammadi from NYU; HGF and TGF β-1 were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: Primer sequences for RT-qPCR.

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques:

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: Total RNA was extracted from ATDC5 cultured in ITS conditions for 0, 7, 14, 21 and 28 days, then reverse transcribed into cDNA and analysed by real-time PCR. The relative abundance of FGF23 (A) was normalized to RPS29 mRNA. Comparison was made by using the ΔΔCt method with the fold value of reference (fold = 1) assigned to D0. *: p < 0.05 vs D0, **: p< 0,01 vs D0. Data are expressed as mean ± SD, n = 4. FGF23 production in media was quantified by ELISA (B), **: p< 0,01 vs D0, ***: p< 0,001, ND: Non Detectable. Total proteins were extracted from ATDC5 (0, 7, 14, 21 or 28 days post-insulin) and 10 μg was subjected to SDS-PAGE with FGF23 (1/200) antibody; ß-actin was used as loading control (C).

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques: Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Comparison, Enzyme-linked Immunosorbent Assay, SDS Page, Control

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: Total RNA was extracted from ATDC5 cultured in ITS conditions for 14 or 28 days and stimulated or not with 100ng/mL mouse rFGF23 for 24h, then reverse transcribed into cDNA and analysed by real-time PCR. The relative abundance at D14 and D28 of FGF23 (A,B), COLX (C,D) and MMP13 (E,F) was normalized to RPS29 mRNA. Comparison was made by using the ΔΔCt method with the fold value of reference (fold = 1) assigned to D0. *: p < 0.05 vs D0, # p<0.05 vs ITS. Data are expressed as mean ± SD, n = 3.

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques: Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Comparison

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: ATDC5 were fixed with 4% PFA, and then stained by Alizarin red for 45min after 21 or 28 days of insulin stimulation with or without 100 ng/mL of mouse rFGF23. Images presented are representative of 5 independent experiments (A). ATDC5 micromasses were cultured for 14 days with insulin stimulation in the presence of 3 mM Pi or 100 ng/mL rFGF23 or both. Then they were fixed with 4% PFA and stained by Alizarin red for 45min. Images presented are representative of 3 independent experiments (B). Total RNA was extracted from ATDC5 cultured in ITS conditions for 14 days, then reverse transcribed into cDNA and analysed by real-time PCR and compared to ITS. The relative abundance of COL X, MMP13 and FGF23 were normalized to RPS29 mRNA (C, D, E). Comparison was made by using the ΔΔCt method with the fold value of reference (fold = 1) assigned to ITS. *: p < 0.05 vs ITS, #: p < 0.05 vs ITS + Pi and £: p < 0.05 vs ITS + rFGF23. Data are expressed as mean ± SD, n = 3.

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques: Staining, Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Comparison

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: Effect of specific inhibition of FGFRs activation during ATDC5 differentiation process (Day 14).

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques: Inhibition, Activation Assay, Expressing

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: Effect of specific inhibition of FGFRs activation during ATDC5 differentiation process (Day 28).

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques: Inhibition, Activation Assay, Expressing

Journal: PLoS ONE

Article Title: Fibroblast-growth factor 23 promotes terminal differentiation of ATDC5 cells

doi: 10.1371/journal.pone.0174969

Figure Lengend Snippet: Total RNA was extracted from ATDC5 cultured in ITS conditions for 28 days pre-treated or not with 1 μM PD173074 one hour before stimulation with 100 ng/ml of rFGF23 (24h for RNA or 48h for proteins), then reverse transcribed into cDNA and analysed by real-time PCR. The relative abundance of FGF23 (A), MMP13 (B) and COL X (C) was normalized to RPS29 mRNA. Comparison was made by using the ΔΔCt method with the fold value of reference (fold = 1) assigned to D0. *: p < 0.05 vs D0, # p< 0.05 vs ITS, £: p< 0.01 vs rFGF23. Data are expressed as mean ± SD, n = 3.

Article Snippet: When required, after serum deprivation, cells were stimulated by 100 ng/mL of recombinant mouse FGF23 (rFGF23, R&D Systems, UK) for 24h at 7, 14, 21 or 28 days of culture and treated with 0.5 or 1 μM of PD173074 (Sigma-Aldrich, France), a pan-FGFR inhibitor, during the culture.

Techniques: Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Comparison

Journal: Reproduction

Article Title: Culture conditions antagonize lineage-promoting signaling in the mouse blastocyst

doi: 10.1530/rep-20-0107

Figure Lengend Snippet: Figure 1 BSA antagonizes signaling by exogenous FGF4. (A) Experimental design for embryos shown and analyzed in this figure. Red = NANOG indicative of epiblast cell fate, green = SOX17 indicative of primitive endoderm cell fate. (B) Immunostaining for SOX17 to identify primitive endoderm (PE) cells and NANOG to identify epiblast (EPI) cells in embryos cultured for 66 hours starting from two-cell stage in KSOM with 1 mg/mL BSA or without BSA (Millipore-Sigma MR-121-D and MR-107-D, respectively), but with 1 mg/mL added PVA (Sigma 360627). n = number of embryos examined. P = Student’s t-test. n.s. = not significant (P > 0.2). (C) Quantification of the total inner cell mass (ICM) and trophectoderm (TE) cells in embryos cultured in indicated conditions and proportion of ICM cells contributing to PE and EPI in these embryos. Each symbol represents a single embryo, column = mean, and error bars = standard deviations. P = Student’s t-test. n.s. = not significant (P > 0.2). (D) Immunostaining for SOX17 and NANOG in embryos cultured in KSOM + 150 ng/mL FGF4 (R&D Systems 235-F4-025) and 1 µg/mL heparin (Sigma H3149) in the presence or absence of 1 mg/mL BSA. Each symbol represents a single embryo, column = mean, and error bars = s.d. P = Student’s t-test. n.s. = not significant (P > 0.2). (E) Quantification of lineage specification in embryos cultured in conditions indicated. Each symbol represents a single embryo, column = mean, and error bars = s.d. (F) Response of embryos cultured in KSOM without 1 mg/mL BSA to low dose of exogenous FGF4 and 1 µg/mL heparin. Each symbol represents a single embryo, column = mean, and error bars = s.d. P = Student’s t-test.

Article Snippet: = not significant (P > 0.2). (D) Immunostaining for SOX17 and NANOG in embryos cultured in KSOM + 150 ng/mL FGF4 (R&D Systems 235-F4-025) and 1 μg/mL heparin (Sigma H3149) in the presence or absence of 1 mg/mL BSA.

Techniques: Immunostaining, Cell Culture