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: Molecular medicine reports

Article Title: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration.

doi: 10.3892/mmr.2016.4942

Figure Lengend Snippet: Figure 2. Expression patterns of FGF21 and β‑klotho in skin following wounding. (A) Reverse‑transcription polymerase chain reaction analysis was used to analyze the expression of FGF21 and β‑klotho prior to and 3 h after wounding. Expression levels of (B) FGF21 and (C) β‑klotho were normalized against those of GAPDH. The experiments were repeated at least three times and values are expressed as the mean ± standard error of three replicates. ***P<0.001 vs. Con. Con, control.

Article Snippet: Subsequently, cells were cultured for 72 h with addition of 0.8% methanol every 24 h. Proteins were separated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Beyotime Institute of Biotechnology, Shanghai, China) and western blot analysis was performed using mouse anti-FGF21 monoclonal antibody (1:1,000; MAB25371; R&D Systems, Inc., Minneapolis, MN, USA) (18).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Molecular medicine reports

Article Title: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration.

doi: 10.3892/mmr.2016.4942

Figure Lengend Snippet: Figure 1. FGF21 expression patterns in a variety of mouse tissues. (A) Expression patterns of FGF21 were analyzed by reverse‑transcription polymerase chain reaction analysis. (B) FGF21 levels in A were normal ized against those of GAPDH. The experiments were repeated at least three times and values are expressed as the mean ± standard deviation of three replicates. **P<0.01; ***P<0.001 vs. heart FGF21 levels.

Article Snippet: Subsequently, cells were cultured for 72 h with addition of 0.8% methanol every 24 h. Proteins were separated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Beyotime Institute of Biotechnology, Shanghai, China) and western blot analysis was performed using mouse anti-FGF21 monoclonal antibody (1:1,000; MAB25371; R&D Systems, Inc., Minneapolis, MN, USA) (18).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

Journal: Molecular medicine reports

Article Title: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration.

doi: 10.3892/mmr.2016.4942

Figure Lengend Snippet: Figure 4. Polymerase chain reaction analysis confirmed that the FGF21 frag ment was successfully integrated into SMD1168 colonies. Lanes 1‑10 are colonies selected from a minimal dextrose plate. Yeast cells transfected with empty vector was used as a negative control. M, marker.

Article Snippet: Subsequently, cells were cultured for 72 h with addition of 0.8% methanol every 24 h. Proteins were separated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Beyotime Institute of Biotechnology, Shanghai, China) and western blot analysis was performed using mouse anti-FGF21 monoclonal antibody (1:1,000; MAB25371; R&D Systems, Inc., Minneapolis, MN, USA) (18).

Techniques: Polymerase Chain Reaction, Transfection, Plasmid Preparation, Negative Control, Marker

Journal: Molecular medicine reports

Article Title: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration.

doi: 10.3892/mmr.2016.4942

Figure Lengend Snippet: Figure 3. Gene synthesis of FGF21. PCR products of the gene FGF21. Lanes: M, 2000 DNA marker; 1, PCR reaction with primer pair P1/RP1; 2, PCR reaction with primer pair P2/RP2 using the PCR product from the previous cycle as a template; 3‑7, PCR reactions with primer pairs P3/RP3‑P7/RP7, respectively, using the PCR product from the previous round as a template. PCR, polymerase chain reaction.

Article Snippet: Subsequently, cells were cultured for 72 h with addition of 0.8% methanol every 24 h. Proteins were separated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Beyotime Institute of Biotechnology, Shanghai, China) and western blot analysis was performed using mouse anti-FGF21 monoclonal antibody (1:1,000; MAB25371; R&D Systems, Inc., Minneapolis, MN, USA) (18).

Techniques: Marker, Polymerase Chain Reaction

Journal: Molecular medicine reports

Article Title: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration.

doi: 10.3892/mmr.2016.4942

Figure Lengend Snippet: Figure 6. Effects of exogenous FGF21 treatment on the activation of cell migration and JNK phosphorylation levels. (A) Wound healing assay was performed to analyze the effects of FGF21 (100 ng/ml) in human fibroblasts under low‑glucose (5.5 mM) conditions. Scale bar, 0.05 µm. Magnification, 40x. (B) The cell migra tion distance was quantified from A. (C and D) Following 6 h culture, 100 ng/ml FGF21 was added to the culture medium and gently shaken. Phosphorylation levels of JNK were analyzed after 30 min of FGF21 stimulation. All experiments were performed after incubation with 5 µg/ml mitomycin‑C, an inhibitor of cell proliferation, for one day. Images were captured using an ImageQuant LAS 4000. Values are expressed as the mean ± standard error (n=10 for B and n=3 for D). *P<0.05 vs. control. p‑JNK, phosphorylated c‑Jun N‑terminal kinase; t‑JNK, total c‑Jun N‑terminal kinase; FGF, fibroblast growth factor; Con, control.

Article Snippet: Subsequently, cells were cultured for 72 h with addition of 0.8% methanol every 24 h. Proteins were separated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Beyotime Institute of Biotechnology, Shanghai, China) and western blot analysis was performed using mouse anti-FGF21 monoclonal antibody (1:1,000; MAB25371; R&D Systems, Inc., Minneapolis, MN, USA) (18).

Techniques: Activation Assay, Migration, Phospho-proteomics, Wound Healing Assay, Incubation, Control

Journal: Molecular medicine reports

Article Title: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration.

doi: 10.3892/mmr.2016.4942

Figure Lengend Snippet: Figure 5. Protein expression and purification of FGF21. (A) SDS‑PAGE analysis of supernatant of engineered P. pastoris cells and purified FGF21 protein. Lanes: 1, marker; 2, supernatant without transfection with the FGF21 overexpression vector; 3, supernatant after transfection; 4, purified protein. (B) Purified FGF21 protein was analyzed by western blotting using an Epson Perfection V700 photo scanner. Lanes: 1, protein in the supernatant after transfection; 2, purified protein. (C) The purity of recombinant FGF21 was analyzed by high‑performance liquid chromatography. FGF, fibroblast growth factor.

Article Snippet: Subsequently, cells were cultured for 72 h with addition of 0.8% methanol every 24 h. Proteins were separated by 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Beyotime Institute of Biotechnology, Shanghai, China) and western blot analysis was performed using mouse anti-FGF21 monoclonal antibody (1:1,000; MAB25371; R&D Systems, Inc., Minneapolis, MN, USA) (18).

Techniques: Expressing, Purification, Marker, Transfection, Over Expression, Plasmid Preparation, Western Blot, Recombinant, High Performance Liquid Chromatography

Journal: medRxiv

Article Title: Physiological and molecular characterization of individuals carrying a diabetogenic mtDNA mutation establishes a mitochondrial basis for insulin resistance in humans

doi: 10.64898/2025.12.17.25342274

Figure Lengend Snippet: (A-D) Time course of venous and arterial plasma GDF15 levels during the hyperinsulinemic-euglycemic (HE) clamp. (B) GDF15 levels in venous plasma samples obtained before (Basal) and during the steady-state period of HE clamp (Insulin). (C) Time course of the venous-arterial difference in plasma GDF15 levels during the HE clamp. (D) Venous-arterial difference in plasma GDF15 before (Basal) and during the steady-state period of the HE clamp (Insulin). (E) Time course of venous and arterial plasma FGF21 levels during the HE clamp. n = 29 (15 in m.3243A>G, 14 in Controls). (F) FGF21 levels in venous plasma samples obtained before (Basal) and during the steady-state period of the HE clamp (Insulin). n = 29 (15 in m.3243A>G, 14 in Controls). (G) Time course of the venous-arterial difference in plasma FGF21 levels during the HE clamp. n = 29 (15 in m.3243A>G, 14 in Controls). (H) Venous-arterial difference in plasma FGF21 before (Basal) and during the steady-state period of the HE clamp (Insulin). n = 29 (15 in m.3243A>G, 14 in Controls). Linear mixed models were used to estimate within– and between-group differences [(B), (D), (F), and (H)]. Data are presented as observed individual values (with lines connecting individually matched participants) and estimated means ± 95% confidence limits, unless otherwise stated. For m.3243A>G carriers, individual datapoints are color-shaded to indicate muscle mtDNA heteroplasmy (light red = low, dark red = high). *Different from Basal ( P < 0.05). † Different from zero ( P < 0.05). n = 30 unless otherwise stated.

Article Snippet: Arterial and venous plasma samples were also analyzed for GDF15 and FGF21 using the automated immunoassay platform Ella (ProteinSimple, San Jose, California, USA) with Simple Plex assays for human GDF15 and FGF21 detection, according to the manufacturer’s instructions.

Techniques: Clinical Proteomics

Journal: Frontiers in bioscience (Landmark edition)

Article Title: Determining the Role of SGLT2 Inhibition with Dapagliflozin in the Development of Diabetic Retinopathy.

doi: 10.31083/j.fbl2712321

Figure Lengend Snippet: Fig. 10. Treatment with DAPA increased the beneficial growth factor FGF21 in diabetic Akita and Akimba mice. Graphs show FGF21 levels after 8 weeks of treatment with DAPA or vehicle in (A) Akita and (B) Akimba mice. *p ≤0.05; data represented as mean

Article Snippet: Serum was analyzed for Fibroblast Growth Factor 21 (FGF21) using the Duoset human FGF21 enzyme-linked immunosorbent assay (ELISA) kit (Cat# DY2539, R&D systems, Inc., Minneapolis, MN, USA) according to the manufacturer’s respective instructions.

Techniques:

Journal: PLoS ONE

Article Title: FGF19 Regulates Cell Proliferation, Glucose and Bile Acid Metabolism via FGFR4-Dependent and Independent Pathways

doi: 10.1371/journal.pone.0017868

Figure Lengend Snippet: (A) GAL-Elk1 luciferase assay in rat L6 cells. L6 cells were cotransfected with expression vectors for KLB and the indicated FGFR together with GAL-Elk1, SV40-renilla Luciferase, and Gal-responsive firefly luciferase reporter. Transfected cells were incubated with media containing increasing concentrations of FGF19 (○) or FGF21(▴) for 6 hours before luciferase assays. Transcriptional activation was assessed by the relative firefly luciferase activity normalized by renilla luciferase activity and expressed as relative luciferase unit (RLU). (B) Drawings (to scale) of FGF19 (top), FGF21 (bottom), and various chimeric proteins with amino acid composition at left. Based on the results of repeated GAL-Elk1 assays such as shown in (C), each chimera was classified into class (I), (II) or (III) as indicated at right (see text). Chimeras which did not exhibit an equivalent FGFR1c activity to FGF21 or FGF19 when conditioned medium was used were not shown here. (C) Representative results of GAL-Elk-1 assay for chimeras shown in (B). L6 cells were cotransfected with expression vectors for KLB and/or FGFR as indicated at right. Each FGF construct was expressed in transiently transfected 293 cells and the conditioned medium was used in the assay. The results are shown as a fold induction over control media conditioned with mock transfected cells. (D) Similar to (A). Purified FGF19 (○) and FGF19v (▾), (the construct #4 in (B) and (C)), were tested for FGFR activation in the presence or absence of KLB coexpression as indicated. (E) Solid phase binding assay of FGF19 and FGF19v to FGFR4 fused to Fc fragment was tested as described in the method section. Schematic diagram for the experiments is shown at right. Bold Y indicates antibody against FGF19 (black) or Fc fragment (gray). HRP: horseradish peroxidase. (F) A control ELISA experiment to show that anti-FGF19 antibody used in (E) recognize FGF19 and FGF19v at indistinguishable affinity. Schematic diagram for the experiments is shown at right.

Article Snippet: For some experiments, E. coli derived FGF21 (2539-FG/CF, R&D systems) was used.

Techniques: Luciferase, Expressing, Transfection, Incubation, Activation Assay, Activity Assay, Construct, Control, Purification, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: FGF19 Regulates Cell Proliferation, Glucose and Bile Acid Metabolism via FGFR4-Dependent and Independent Pathways

doi: 10.1371/journal.pone.0017868

Figure Lengend Snippet: 11-week-old ob/ob mice were subcutaneously implanted with an osmotic pump to infuse 1 ng/hr FGF protein (0.4 mg/kg/day) or PBS control (N = 7). (A) Changes in body weight and random fed blood glucose level. The osmotic pump was implanted on day 0. (B) Blood glucose levels at random fed condition and after overnight fast. (C) Serum non-esterified fatty acids (NEFA) levels on day 8. (D) Glucose tolerance test conducted on day 6. Mice were overnight fasted and i.p. injected with 1 g/kg glucose at t = 0. (E) Organ/body weight ratio on day 8. (F) qPCR gene expression profiles on indicated organs. p values: *<0.05, **<0.005, ***<0.0005 (vs PBS control), ##p<0.005 (FGF21 vs FGF19v).

Article Snippet: For some experiments, E. coli derived FGF21 (2539-FG/CF, R&D systems) was used.

Techniques: Control, Injection, Gene Expression

Journal: PLoS ONE

Article Title: FGF19 Regulates Cell Proliferation, Glucose and Bile Acid Metabolism via FGFR4-Dependent and Independent Pathways

doi: 10.1371/journal.pone.0017868

Figure Lengend Snippet: (A) Schematic diagram showing hepatic biosynthetic pathways that convert cholesterol into bile acids. The classical (neutral) pathway (center) is initiated by Cyp7A1, whereas the alternate (acidic) pathway (right) is initiated by Cyp27a1 and Cyp7b1. According to our model, FGF19 suppresses the classical pathway through transcriptional regulation of Cyp7a1 and Cyp8b1, shifting BA synthesis towards production of CDCA or its derivatives. (B) Distinct FGFR/KLB receptor complexes mediate various biological activities of FGF19, FGF19v, and FGF21. In addition to the model depicted, FGF21 and FGF19 can also suppress Cyp7a1 expression acutely in FGFR4-independent manner .

Article Snippet: For some experiments, E. coli derived FGF21 (2539-FG/CF, R&D systems) was used.

Techniques: Expressing