Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 2 | Activating effects of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of PD173074 (FGFR1 inhibitor, 50 nmol L−1) or palmitic acid (PA, 500 µmol L−1), on FGF21 signaling in HepG2 human hepatocytes. Coffee by-product’s bioactive compounds interacted in silico with the subunits of the FGF21 receptor (FGFR) (A), exhibiting different binding (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: In Silico, Binding Assay

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 4 | Role of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (PA, 500 µmol L−1) on lipo/cytotoxicity and inflammation markers in HepG2 human hepatocytes. Coffee by-products’ bioactive compounds preserved cell viability (A) and diminished lactate dehydrogenase (LDH) release (B). PA-triggered tumor necrosis factor α (TNF-α) (C), interleukin (IL)-6 (D), and IL-1β (E) release and nitric oxide synthase (NOS) activity (F) were reduced. Hierarchical cluster analysis and heat map [from the lowest (■red) to the highest (■green) value for each parameter] (G) and an integrative diagram illustrating the effects of the bioactive compounds from coffee by-products on lipo/cytotoxicity and inflammation (H). The results are expressed as mean ± SD (n = 3). Bars with different letters significantly (p < 0.05) differ according to ANOVA and Tukey’s multiple range test. NT, non-treated cells; CAF, caffeine; CGA, chlorogenic acid; CA, caffeic acid; PCA, protocatechuic acid; GA, gallic acid; KMP, kaempferol; FGF21, fibroblast growth factor 21.

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: Activity Assay

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 5 | Protective effects of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (PA, 500 µmol L−1) against oxidative stress in HepG2 human hepatocytes. Coffee by-products’ bioactive compounds diminished the production of reactive oxygen species (ROS) (A) and mitochondrial O•− 2 (B), preserving the mitochondrial membrane potential (19m) (C) in PA-challenged HepG2 cells. The enzymatic activity NADPH oxidase (NOX) (D), superoxide dismutase (SOD) (E), and catalase (F) activities, and nuclear factor (erythroid-derived 2)-like 2 (Nrf2) phosphorylation (G) were regulated, thereby diminishing oxidative stress. Hierarchical cluster analysis and heat map [from the lowest (■red) to the highest (■green) value for each parameter] (H) and an integrative diagram illustrating the effects of the bioactive compounds from coffee by-products on oxidative stress (I). The results are expressed as mean ± SD (n = 3). Bars with different letters significantly (p < 0.05) differ according to ANOVA and Tukey’s multiple range test. NT, non-treated cells; CAF, caffeine; CGA, chlorogenic acid; CA, caffeic acid; PCA, protocatechuic acid; GA, gallic acid; KMP, kaempferol; FGF21, fibroblast growth factor 21.

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: Preserving, Membrane, Activity Assay, Derivative Assay, Phospho-proteomics

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 6 | Regulative role of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (500 µmol L−1) on the mitochondrial bioenergetics of HepG2 human hepatocytes. Coffee (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques:

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 7 | Modulatory effects of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (500 µmol L−1) on lipid metabolism in HepG2 human hepatocytes. Palmitic acid-treated (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques:

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 8 | Regulatory role of standard solutions of the bioactive compounds from coffee by-products (50 µmol L−1), aqueous extracts (CSE and CHE, 100 µg mL−1), and FGF21 (20 nmol L−1), in the presence of palmitic acid (500 µmol L−1) on glucose metabolism in HepG2 human hepatocytes. Hepatocytes exhibited a modulation on glucose uptake (A), glucokinase (GK) activity (B), glucose production (C), phosphoenolpyruvate carboxykinase (PEPCK) activity (D), insulin receptor (Continued)

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques: Activity Assay

Journal: Frontiers in nutrition

Article Title: Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro .

doi: 10.3389/fnut.2022.866233

Figure Lengend Snippet: FIGURE 9 | Integrative hierarchical cluster analysis and heat map (from the lowest (■red) to the highest (■green) value for each parameter) unifying all parameters demonstrates that chlorogenic and protocatechuic acids are the bioactive compounds exhibiting the highest NAFLD-protecting effects (A). Diagram illustrating the molecular mechanisms from the effects of the bioactive compounds in coffee by-products on hepatic FGF21 signaling, oxidative stress, mitochondrial bioenergetics, and energy metabolism (B). NT, non-treated cells; PA, palmitic acid; CAF, caffeine; CGA, chlorogenic acid; CA, caffeic acid; PCA, protocatechuic acid; GA, gallic acid; KMP, kaempferol; FGF21, fibroblast growth factor 21.

Article Snippet: Recombinant human FGF21 was obtained from R&D Systems (Minneapolis, MN, USA), whereas the FGFR Frontiers in Nutrition | www.frontiersin.org 2 March 2022 | Volume 9 | Article 866233 inhibition, PD173074, was purchased from AdoQ Bioscience (Irvine, CA, USA).

Techniques:

Journal: Cell Metabolism

Article Title: GDF15 Provides an Endocrine Signal of Nutritional Stress in Mice and Humans

doi: 10.1016/j.cmet.2018.12.016

Figure Lengend Snippet: GDF15 Expression Is Regulated by the Cellular ISR Pathway (A and C) GDF15 mRNA expression (A) and immunoblot analysis (C) of ISR components in wild-type (WT) mouse embryonic fibroblasts (MEFs) treated with vehicle control (Con), cobalt chloride (CoCl2, 625 μM), thapsigargin (Tg, 1 μM), tunicamycin (Tn 5 μg/mL), or L-Histidinol (His, 1 mM) for 6 h. (B) GDF15 mRNA expression in human cell lines (HeLA, HuH7, and A549) treated with Tn (5 μg/mL) for 6 h. (D) GDF15 mRNA expression in WT MEFs pre-treated for 1 h either with the PERK inhibitor GSK2606414 (GSK, 200 nM) or eIF2α inhibitor ISRIB (ISR, 100 nM), then co-treated with Tn (5 μg/mL) for a further 6 h. (E–G) GDF15 mRNA expression (E) in EIF2α Ser51 (SS) or phospho mutant (AA) MEFs or (F) in ATF4 wild-type (WT) or ATF4 knockout (KO) MEFs and (G) in control siRNA and CHOP siRNA transfected WT MEFs treated with Tn (5 μg/mL) for 6 h. (H) Diagram outlining pathway by which GDF15 and FGF21 expression is regulated by TN. mRNA expression is presented as fold expression relative to its respective control treatment for each cell type (set at 1) or TN-treated samples (set as 100) with normalization to HPRT gene expression in MEFs and GAPDH in human cells. Data are expressed as mean ± SD from at least three independent experiments. ∗∗∗ p < 0.001 versus control (con) for (A) and (B), and versus TN stimulated for (D)–(G) by two-tailed Student’s t test. Blots shown are representative of three independent experiments with Calnexin used as a loading control. See also Figures S3 and .

Article Snippet: Mouse FGF21 was analyzed by FGF21 Quantakine ELISA (R&D Systems) following the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Control, Mutagenesis, Knock-Out, Transfection, Gene Expression, Two Tailed Test

Journal: Cell Metabolism

Article Title: GDF15 Provides an Endocrine Signal of Nutritional Stress in Mice and Humans

doi: 10.1016/j.cmet.2018.12.016

Figure Lengend Snippet:

Article Snippet: Mouse FGF21 was analyzed by FGF21 Quantakine ELISA (R&D Systems) following the manufacturer’s instructions.

Techniques: Recombinant, SYBR Green Assay, Protease Inhibitor, Reverse Transcription, Western Blot, Injection, Enzyme-linked Immunosorbent Assay, Control, TaqMan Assay, Software, Sterility, Electrophoresis, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Novel Insights into the Cardio-Protective Effects of FGF21 in Lean and Obese Rat Hearts

doi: 10.1371/journal.pone.0087102

Figure Lengend Snippet: FGF21 mRNA [ Fig. 2A1 ] and protein expression [ Fig. 2A2 ]; βKlotho mRNA [ Fig. 2A3 ] and protein expression [ Fig. 2A4 ] in isolated rat cardiomyocytes [A] and rat heart [B]. Fig . 1B:Immunocyto/histochemistry and confocal analysis of FGF21 protein in isolated adult rat cardiomyocytes [ B1 ] and rat heart [ B2 ]. MW.- Molecular Weight Marker for PCR products; BP.- Base Pairs; kDa.-kilo daltons. Fig. 2C -( a ): Trace recordings of left ventricular developed pressure [LVDP (mmHg)] and left ventricular contractility (dp/dt) in control (saline treated) groups; and Fig. 2C -( b ):[LVDP-mmHg] and dp/dt ratio in FGF21 treated groups - following 30 mins of global ischemia and 120 mins reperfusion. Fig. 2D: Rate pressure product [RPP (mmHg/min)] during global ischemia and reperfusion with or without FGF21 treatment [** P <0.01 vs. control]. Fig. 2E : Graphical representation of infarct area (%) in rat hearts treated with or without FGF21. Data shown are means ± SEM (n = 6, in triplicates). ***P<0.001; **P<0.01 vs. control.

Article Snippet: The drugs used for the study were: rat recombinant FGF21 [R&D systems, UK], wortmannin (PI3K inhibitor), U1026 (MAPK inhibitor), Compound C (AMPK inhibitor), TO-901317 [ROR-α (Retinoic acid receptor-related receptor alpha) inhibitor]; TO-901317 (was initially intended as a therapeutic Liver X receptor (LXR) agonist in dementia , however recently, work by Uebanso T et al, have demonstrated its role in the regulation of FGF21 expression ) and collagenase (type I+protease type XIV).

Techniques: Expressing, Isolation, Molecular Weight, Marker, Control, Saline

Journal: PLoS ONE

Article Title: Novel Insights into the Cardio-Protective Effects of FGF21 in Lean and Obese Rat Hearts

doi: 10.1371/journal.pone.0087102

Figure Lengend Snippet: Fig. 3A1: Cardiomyocytes treated with FGF21 (100 nM) for 5–30 minutes. Phosphorylated ERK 1/2 [Fig. 3A1]; Akt [Fig. 3A2] and AMPK [Fig. 3A3] proteins are represented in relation to total proteins, expressed as fold increase over basal. ***P<0.001, **P<0.01, *P<0.05 vs. basal, n = 6 per group. Fig. 3B1: RPP with ischemia/reperfusion and FGF21 treatment following pre-incubation with inhibitors (TO-901317; wortmanin; Compound C or U0126) a P<0.05, b P<0.01 vs. FGF21 only treatment, n = 6 per group. Fig. 3B2: Infarcted area (%) following ischemia/reperfusion and FGF21 treatment following pre-incubation with inhibitors (TO-901317; wortmanin; Compound C or U0126) **P<0.01, *P<0.05 vs. FGF21 treated, n = 6 per group.

Article Snippet: The drugs used for the study were: rat recombinant FGF21 [R&D systems, UK], wortmannin (PI3K inhibitor), U1026 (MAPK inhibitor), Compound C (AMPK inhibitor), TO-901317 [ROR-α (Retinoic acid receptor-related receptor alpha) inhibitor]; TO-901317 (was initially intended as a therapeutic Liver X receptor (LXR) agonist in dementia , however recently, work by Uebanso T et al, have demonstrated its role in the regulation of FGF21 expression ) and collagenase (type I+protease type XIV).

Techniques: Incubation

Journal: PLoS ONE

Article Title: Novel Insights into the Cardio-Protective Effects of FGF21 in Lean and Obese Rat Hearts

doi: 10.1371/journal.pone.0087102

Figure Lengend Snippet: Fig. 4A :FGF21 mRNA expression levels in cardiomyocytes following FGF21 (100 nM) treatment with or without pathway inhibitors [(U0126; wort (wortmanin); Comp C (Compound C) or TO (TO-901317)] (normalised to GAPDH and expressed as fold changes over basal). Fig. 4B : Graphical analysis of FGF21 protein levels following FGF21 treatment. Fig. 4C : Graphical representation of FGF21 ELISA measurements in the conditioned media following FGF21 treatment. Fig. 4D : Graphical representation of FGF21 ELISA measurements of rat heart Langendorff exudates following global ischemia for 5–30 mins and 120 mins of reperfusion; with or without prior FGF21 (100 nM) infusion. Data shown are means ± SEM of triplicates. The values represented are relative to basal. *** P <0.001; ** P <0.01; * P <0.05 vs. FGF21 only treated, a P <0.001 vs. basal, n = 6 per group.

Article Snippet: The drugs used for the study were: rat recombinant FGF21 [R&D systems, UK], wortmannin (PI3K inhibitor), U1026 (MAPK inhibitor), Compound C (AMPK inhibitor), TO-901317 [ROR-α (Retinoic acid receptor-related receptor alpha) inhibitor]; TO-901317 (was initially intended as a therapeutic Liver X receptor (LXR) agonist in dementia , however recently, work by Uebanso T et al, have demonstrated its role in the regulation of FGF21 expression ) and collagenase (type I+protease type XIV).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Novel Insights into the Cardio-Protective Effects of FGF21 in Lean and Obese Rat Hearts

doi: 10.1371/journal.pone.0087102

Figure Lengend Snippet: Using rat heart Langendorff model and inducing global ischemia for 30[ Fig. 5A1 ], protein [ Fig. 5A2 ], and secretion of FGF21 in Langendorff coronary exudates [ Fig. 5A3 ] were measured. Similarly, changes in FGFR1 mRNA expressions [ Fig. 5A4 ] were measured. Data shown are means ± SEM of triplicates. The values represented are relative to basal. *** P <0.001; ** P <0.01vs. control. n = 6 per group. FGF21 mRNA [ Fig. 5B1 ], protein [ Fig. 5B2 ], FGF21 secretion (in Langendorff coronary exudates) [ Fig. 5B3 ] and FGFR1 mRNA [ Fig. 5B4 ] expressions were measured in obese and lean rat hearts. Graphical representation of a key signalling component of FGF21/FGFR1; βKlotho protein level in obese and lean hearts [ P <0.05 vs.lean; Figure 5C 1 ]. Graphical representation of ERK 1/2 [ Fig. 5C2 ], Akt [ Fig. 5C3 ] and AMPK [ Fig. 5C4 ] phosphorylation levels in lean and obese hearts with FGF21 (100 nM) pre-infusion. Data shown are means ± SEM of triplicates. The values represented are relative to basal. * P <0.05, ** P <0.01 vs. lean control, NS -non-significant; n = 6 per group.

Article Snippet: The drugs used for the study were: rat recombinant FGF21 [R&D systems, UK], wortmannin (PI3K inhibitor), U1026 (MAPK inhibitor), Compound C (AMPK inhibitor), TO-901317 [ROR-α (Retinoic acid receptor-related receptor alpha) inhibitor]; TO-901317 (was initially intended as a therapeutic Liver X receptor (LXR) agonist in dementia , however recently, work by Uebanso T et al, have demonstrated its role in the regulation of FGF21 expression ) and collagenase (type I+protease type XIV).

Techniques: Control, Phospho-proteomics

Journal: PLoS ONE

Article Title: Novel Insights into the Cardio-Protective Effects of FGF21 in Lean and Obese Rat Hearts

doi: 10.1371/journal.pone.0087102

Figure Lengend Snippet: LVDP (mmHg) and left ventricular contractility (dp/dt) in obese control (saline treated) groups [ Fig. 6A1 ] and lean control (saline treated) groups [ Fig. 6A2 ]; following global ischemia and reperfusion. Graphical representation of RPP (mmHg/min)] during global ischemia and reperfusion in obese and lean control rat hearts [ Fig. 6B ]. Graphical representation of the infarcted area (%) in obese and lean control rat hearts following global ischemia and reperfusion [ Fig. 6C ]. Graphical representation of FGF21 levels in obese and lean rat heart coronary effluents following global ischemia and reperfusion [ Fig. 6D ]. Data shown are means ± SEM of triplicates. The values represented are relative to basal. *** P <0.001 vs. t [0] time point, n = 6 per group.

Article Snippet: The drugs used for the study were: rat recombinant FGF21 [R&D systems, UK], wortmannin (PI3K inhibitor), U1026 (MAPK inhibitor), Compound C (AMPK inhibitor), TO-901317 [ROR-α (Retinoic acid receptor-related receptor alpha) inhibitor]; TO-901317 (was initially intended as a therapeutic Liver X receptor (LXR) agonist in dementia , however recently, work by Uebanso T et al, have demonstrated its role in the regulation of FGF21 expression ) and collagenase (type I+protease type XIV).

Techniques: Control, Saline

Journal: PLoS ONE

Article Title: Novel Insights into the Cardio-Protective Effects of FGF21 in Lean and Obese Rat Hearts

doi: 10.1371/journal.pone.0087102

Figure Lengend Snippet: LVDP (mmHg) and left ventricular contractility (dp/dt) in obese control (saline treated) groups [ Fig. 7A1 ], obese (FGF21 treated) groups [ Fig. 7A2 ] and lean (FGF21 treated) groups [ Fig. 7A3 ]; following global ischemia and reperfusion. Graphical representation of the infarcted area (%) in obese control, obese and lean FGF21 (100 nM) treated rat hearts following global ischemia and reperfusion [ Fig. 7B ]. Graphical representation of FGF21 levels in obese and lean FGF21 (100nM) pre-treated rat heart coronary effluents following global ischemia and reperfusion [ Fig 6C ] . Data shown are means ± SEM of triplicates. The values represented are relative to basal. *** P < 0.001 vs. t [0] time point, # P < 0.05 vs. lean at t(5); n = 6 per group.

Article Snippet: The drugs used for the study were: rat recombinant FGF21 [R&D systems, UK], wortmannin (PI3K inhibitor), U1026 (MAPK inhibitor), Compound C (AMPK inhibitor), TO-901317 [ROR-α (Retinoic acid receptor-related receptor alpha) inhibitor]; TO-901317 (was initially intended as a therapeutic Liver X receptor (LXR) agonist in dementia , however recently, work by Uebanso T et al, have demonstrated its role in the regulation of FGF21 expression ) and collagenase (type I+protease type XIV).

Techniques: Control, Saline

Journal: Journal of Clinical Investigation

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

doi: 10.1172/jci94337

Figure Lengend Snippet: Figure 3. FGF21 secreted from pancreas induces OPC proliferation. (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 the role of FGFs, we used recombinant mouse FGF15 (Abcam), recombinant mouse FGF21 (R&D Systems), and recombinant mouse FGF23 (R&D Systems).

Techniques: Immunolabeling, Immunohistochemistry, BrdU Incorporation Assay, Knockdown

Journal: Journal of Clinical Investigation

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

doi: 10.1172/jci94337

Figure Lengend Snippet: Figure 6. FGF21 promotes human OPC proliferation. (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 the role of FGFs, we used recombinant mouse FGF15 (Abcam), recombinant mouse FGF21 (R&D Systems), and recombinant mouse FGF23 (R&D Systems).

Techniques: Expressing, BrdU Incorporation Assay, Recombinant, Control

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