Journal: PLoS ONE

Article Title: Fibroblast Growth Factor 21 (FGF21) Protects against High Fat Diet Induced Inflammation and Islet Hyperplasia in Pancreas

doi: 10.1371/journal.pone.0148252

Figure Lengend Snippet: FGF21 is regulated by the nutritional status in fed, fasted and diet-induced obesity (DIO). Fasting reduced FGF21 mRNA in whole pancreas (A). β-klotho, the co-receptor of FGF21, is expressed in the pancreas and is inversely regulated by fasting (B). FGF21 protein was only detected in pancreas compared to liver and inguinal white adipose tissue (IWAT) (C). Pancreas FGF21 protein level was reduced by fasting in both chow fed and high fat diet fed conditions (D, E). Separation of pancreatic fractions to evaluate the individual contribution of islets and acinar tissue on FGF21 expression show that acinar pancreas contributes to the majority of FGF21 levels while islet cells have very low FGF21 levels and are not nutritionally regulated (F). Acinar expression of FGF21 is regulated with fasting (F, G). n = 6 per group. Each western blot lane represents an individual animal.

Article Snippet: Antibodies were purchased from commercial vendors: FGF21 (AF3057; R&D systems, Minneapolis, MN); Insulin (ab7842; Abcam, Cambridge, MA); β-actin (ab8226; Abcam); α -tubulin (ab7291; Abcam); pERK1/2 (4370; Cell Signaling Technology, Denver, MA); ERK1/2 (9107; Cell Signaling Technology); glucagon (2654; Sigma Aldrich, St. Louis, MO); somatostatin (sc7819; Santa Cruz, Dallas, TX); CD3 (MA1-90582; Thermo Scientific, Waltham, MA).

Techniques: Expressing, Western Blot

Journal: PLoS ONE

Article Title: Fibroblast Growth Factor 21 (FGF21) Protects against High Fat Diet Induced Inflammation and Islet Hyperplasia in Pancreas

doi: 10.1371/journal.pone.0148252

Figure Lengend Snippet: Acute FGF21 exposure induces ERK1/2 signaling events therefore phosphorylation of ERK1/2 (pERK1/2) has been chosen as a marker of FGF21 signaling. Up to 50% of acinar cells show strong nuclear pERK1/2 staining (brown) after FGF21 administration (A i, iii). Islets represent a very unique pattern with pERK1/2 only on the peripheral cells (A ii, iv). Percentage of pERK1/2-labeled acinar cells with saline and FGF21 administration (B). Western blot of pERK1/2 and total ERK1/2 from the same pancreas samples (C). Epididymal white adipose tissue (EWAT) is a positive control for FGF21 signaling (C). S = saline and F = FGF21. Confocal photomicrographs show FGF21-induced pERK1/2 immunoreactivity in islets (D). Absence of pERK1/2 staining (green) after saline treatment (Di; Inset , islet outline is indicated by insulin-immunoreactivity shown in red). FGF21 did not elicit pERK1/2 in insulin (red)-producing β-cells (D ii). Merged images show pERK1/2 co-localization in glucagon-positive α cells (D iii) and somatostatin-positive δ cell (D iv). Split panels show glucagon (iii a ) and pERK (iii b ) labeling or somatostatin (iv a ) and pERK (iv b ) labeling in single cells. A robust and specific pERK1/2 staining was observed in almost 10% of total islet nuclei mostly at the periphery. Outlined region (dashed line) in the inset indicates the region represented in each panel. Representative pERK1/2 labelled cells are indicated by white arrows. n = 4 per group. Scale: 200 μm, A; 20 μm, D.

Article Snippet: Antibodies were purchased from commercial vendors: FGF21 (AF3057; R&D systems, Minneapolis, MN); Insulin (ab7842; Abcam, Cambridge, MA); β-actin (ab8226; Abcam); α -tubulin (ab7291; Abcam); pERK1/2 (4370; Cell Signaling Technology, Denver, MA); ERK1/2 (9107; Cell Signaling Technology); glucagon (2654; Sigma Aldrich, St. Louis, MO); somatostatin (sc7819; Santa Cruz, Dallas, TX); CD3 (MA1-90582; Thermo Scientific, Waltham, MA).

Techniques: Phospho-proteomics, Marker, Staining, Labeling, Saline, Western Blot, Positive Control

Journal: PLoS ONE

Article Title: Fibroblast Growth Factor 21 (FGF21) Protects against High Fat Diet Induced Inflammation and Islet Hyperplasia in Pancreas

doi: 10.1371/journal.pone.0148252

Figure Lengend Snippet: Chronic FGF21 treatment for 3 days by mini osmotic pumps led to the reduction in FGF21 expression in pancreas (A). FGF21 infusion down-regulated insulin but glucagon, somatostatin or amylase expression. While FGF21 infusion did not alter serum glucose levels (B), it reduced the serum levels of pancreatic hormones, insulin (C) and glucagon (D) without affecting the amylase activity (E). n = 10 per group.

Article Snippet: Antibodies were purchased from commercial vendors: FGF21 (AF3057; R&D systems, Minneapolis, MN); Insulin (ab7842; Abcam, Cambridge, MA); β-actin (ab8226; Abcam); α -tubulin (ab7291; Abcam); pERK1/2 (4370; Cell Signaling Technology, Denver, MA); ERK1/2 (9107; Cell Signaling Technology); glucagon (2654; Sigma Aldrich, St. Louis, MO); somatostatin (sc7819; Santa Cruz, Dallas, TX); CD3 (MA1-90582; Thermo Scientific, Waltham, MA).

Techniques: Expressing, Activity Assay

Journal: PLoS ONE

Article Title: Fibroblast Growth Factor 21 (FGF21) Protects against High Fat Diet Induced Inflammation and Islet Hyperplasia in Pancreas

doi: 10.1371/journal.pone.0148252

Figure Lengend Snippet: WT and FGF21 KO mice on chow diet have similar body weight (A), glucose tolerance (B) and insulin secretion in response to glucose (C), despite notable differences in their pancreatic morphology (D i, iii). After consuming high fat diet for 16 weeks, FGF21 KO mice (D iv) had increased islet hyperplasia compared with WT mice (D ii). Islet cell surface area was measured and represented (E). Islet cell proliferation markers were substantially over expressed in obese FGF21 KO animals (F). Islet hyperplasia leads to increased serum insulin (G) in FGF21 KO animals with a slight reduction in blood glucose (H) (n = 7 per group). Scale: 1 mm.

Article Snippet: Antibodies were purchased from commercial vendors: FGF21 (AF3057; R&D systems, Minneapolis, MN); Insulin (ab7842; Abcam, Cambridge, MA); β-actin (ab8226; Abcam); α -tubulin (ab7291; Abcam); pERK1/2 (4370; Cell Signaling Technology, Denver, MA); ERK1/2 (9107; Cell Signaling Technology); glucagon (2654; Sigma Aldrich, St. Louis, MO); somatostatin (sc7819; Santa Cruz, Dallas, TX); CD3 (MA1-90582; Thermo Scientific, Waltham, MA).

Techniques:

Journal: PLoS ONE

Article Title: Fibroblast Growth Factor 21 (FGF21) Protects against High Fat Diet Induced Inflammation and Islet Hyperplasia in Pancreas

doi: 10.1371/journal.pone.0148252

Figure Lengend Snippet: HFD consumption caused severe pancreatic inflammation in FGF21 KO mice (A). H&E staining showing, that compared to WT animals (i), FGF21 KO mice developed severe pancreatic periductal inflammation (outlined area in ii-iii) when consuming a HFD diet for 16 weeks. Lymphocytic nature of inflammatory cells is shown in a higher magnification image (iii). (B) Immunohistochemical analysis for the lymphocytic marker CD3 on the pancreas of WT (i) and FGF21 KO animals (ii-iii) is represented. FGF21 KO showed higher number of CD3, T cell receptor antigen. n = 7 per group. Scale: 1 mm, Ai-ii; 2 mm, Bi-ii; 200 μm, iii.

Article Snippet: Antibodies were purchased from commercial vendors: FGF21 (AF3057; R&D systems, Minneapolis, MN); Insulin (ab7842; Abcam, Cambridge, MA); β-actin (ab8226; Abcam); α -tubulin (ab7291; Abcam); pERK1/2 (4370; Cell Signaling Technology, Denver, MA); ERK1/2 (9107; Cell Signaling Technology); glucagon (2654; Sigma Aldrich, St. Louis, MO); somatostatin (sc7819; Santa Cruz, Dallas, TX); CD3 (MA1-90582; Thermo Scientific, Waltham, MA).

Techniques: Staining, Immunohistochemical staining, Marker

Journal: PLoS ONE

Article Title: Fibroblast Growth Factor 21 (FGF21) Protects against High Fat Diet Induced Inflammation and Islet Hyperplasia in Pancreas

doi: 10.1371/journal.pone.0148252

Figure Lengend Snippet: (A) Representative cytofluorometric dot plots of isolated lymphocytes from WT and FGF21 KO animals consuming high fat diet for 16 weeks. FGF21 KO mice show elevated CD45+ (i), TCRb+ and Thy1+ T lymphocytes (ii) and Foxp3+ Treg cells. CD19+ B lymphocytes were not significantly altered (iii). Corresponding summary data is shown in the right panel. The experiment was repeated twice (n = 4 per group). (B) Gene expression analysis of cytokines is presented. n = 7 per group.

Article Snippet: Antibodies were purchased from commercial vendors: FGF21 (AF3057; R&D systems, Minneapolis, MN); Insulin (ab7842; Abcam, Cambridge, MA); β-actin (ab8226; Abcam); α -tubulin (ab7291; Abcam); pERK1/2 (4370; Cell Signaling Technology, Denver, MA); ERK1/2 (9107; Cell Signaling Technology); glucagon (2654; Sigma Aldrich, St. Louis, MO); somatostatin (sc7819; Santa Cruz, Dallas, TX); CD3 (MA1-90582; Thermo Scientific, Waltham, MA).

Techniques: Isolation, Gene Expression

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: Scientific Reports

Article Title: Fgf21 regulates T-cell development in the neonatal and juvenile thymus

doi: 10.1038/s41598-017-00349-8

Figure Lengend Snippet: Fgf21 mRNA is enriched in mature mTECs. ( A and B ) Expression levels of Fgf21 mRNA in the various tissues from 4-week-old C57BL/6 mice ( A ) and in the thymus of E15.5 and 0-, 4-, 8- and 12-week-old ( B ). Relative Fgf21 mRNA expression was normalised against 18S rRNA expression. ( C and D ) Thymic cell suspensions from 4-week-old C57BL/6 mice were assessed by flow cytometric analysis using anti-CD45 mAb, anti-EpCAM mAb, anti-MHC II (I-A/I-E) mAb, and UEA-1 lectin, and expression levels of Fgf21 mRNA were determined by RT-realtime PCR in the subpopulations of thymic cells from 4-week-old C57BL/6 mice. ( C ) Anti-CD45 and anti-EpCAM staining discriminated CD45 + , TEC (CD45 − EpCAM + ), and non-TEC stromal cell (CD45 − EpCAM − ) subpopulations. ( D ) Gated TECs were subdivided into cTEC hi (UEA-1 − MHC high ), cTEC lo (UEA-1 − MHC low ), mTEC hi (UEA-1 + MHC high ), and mTEC lo (UEA-1 + MHC low ) subpopulations. ( E ) Thymocytes were removed from embryonic thymi by 1.35 mM dGuo treatment for 6 days, and remaining stromal cells were stimulated with 1 μg/ml RANKL to induce the maturation of TECs. Fgf21 and Aire mRNA were increased along with TEC maturation. Graphs represent the mean ± SD; n ≧5 replicates per group from at least 2 independent experiments.

Article Snippet: Medium containing 500 ng/ml of recombinant human FGF21 (R&D Systems) was replaced at 3-day intervals.

Techniques: Expressing, Staining

Journal: Scientific Reports

Article Title: Fgf21 regulates T-cell development in the neonatal and juvenile thymus

doi: 10.1038/s41598-017-00349-8

Figure Lengend Snippet: CD4SP and CD8SP populations are decreased in Fgf21 KO mice. ( A and B ) Thymi and spleens were isolated from 4-week-old WT and Fgf21 KO mice, and their weights were measured. ( C ) Cell numbers of enzymatically digested suspensions from thymi and spleens were counted. ( D ) Thymocytes and splenocytes from WT and Fgf21 KO mice were stained with anti-CD4 and anti-CD8 mAb. Representative flow cytometry plots show CD4/CD8 analysis of thymocytes and splenocytes from 1-week-old WT and Fgf21 KO mice. ( E ) Charts show the percentage of DN, DP, CD8SP, and CD4SP cells from 1- and 4-week-old WT and Fgf21 KO mice. ( F ) Charts show the percent of CD4SP and CD8SP cells from 1- and 4-week-old WT and Fgf21 KO mice. ( G ) Thymocytes from 1-week-old WT and Fgf21 KO mice were defined by TCRβ and CD69 levels and subdivided into 5 subsets (T1; TCRβ − CD69 − , T2; TCRβ int CD69 − , T3; TCRβ int CD69 + , T4; TCRβ hi CD69 + , and T5; TCRβ hi CD69 − ). ( H ) Charts show the percentage of the 5 subsets from 1-week-old WT and Fgf21 KO mice. ( I ) Gated CD4SP and CD8SP cells from 1-week-old WT and Fgf21 KO mice were defined by CD62L and CD24 levels and subdivided into CD24 + CD62L − immature and CD24 − CD62L + mature SP cells. ( J ) Charts show the percentage of CD24 + CD62L − immature and CD24 − CD62L + mature SP subsets from 1-week-old WT and Fgf21 KO mice. All data shown are the mean ± SD from ≧6 mice per genotype from 2 independent experiments. *P < 0.05, *P < 0.01 versus WT mice.

Article Snippet: Medium containing 500 ng/ml of recombinant human FGF21 (R&D Systems) was replaced at 3-day intervals.

Techniques: Isolation, Staining, Flow Cytometry

Journal: Scientific Reports

Article Title: Fgf21 regulates T-cell development in the neonatal and juvenile thymus

doi: 10.1038/s41598-017-00349-8

Figure Lengend Snippet: Thymic stromal cells are not obviously altered in Fgf21 KO mice. ( A ) Enzymatically digested thymic cell suspensions were stained with anti-CD45 mAb and anti-EpCAM mAb, and the TEC (CD45 − EpCAM + ) number was counted from the thymi of E14.5, E15.5, E18.5, and 4-week-old WT and Fgf21 KO mice. ( B ) Immunohistochemistry of WT and Fgf21 KO thymi with anti-Keratin-5 antibody for a medullary TEC marker (left panels) and anti-Ly-51 antibody for a cortical TEC marker (right panels). ( C ) The ratio of Keratin-5-positive thymic medullary regions was measured from immunostained sections. ( D ) Gated TECs of 1-weeks old were subdivided into mTEC hi (Ly51 − MHC high ), mTEC lo (Ly51 − MHC low ), cTEC hi (Ly51 + MHC high ), and cTEC lo (Ly + MHC low ) subpopulations. Graphs represent the percentage of subpopulations. ( E ) The mean fluorescence intensity (MFI) of surface MHCII on the TEC was measured by flow cytometry with anti-MHCII antibody. ( F ) Gated CD45 − thymic stromal cells of 1-weeks old were stained with anti-CD140a and anti-CD31 antibodies. Graphs represent the cellularity of CD45 − CD140a + fibroblasts and CD45 − CD31a + endothelial cells in the thymus of WT and Fgf21 KO mice. ( G ) The expression levels of thymic factors were measured in the thymus of 1-week-old WT and Fgf21 KO mice. Graphs represent the mean ± SD; n ≧ 5 replicates per group from at least 3 independent experiments.

Article Snippet: Medium containing 500 ng/ml of recombinant human FGF21 (R&D Systems) was replaced at 3-day intervals.

Techniques: Staining, Immunohistochemistry, Marker, Fluorescence, Flow Cytometry, Expressing

Journal: Scientific Reports

Article Title: Fgf21 regulates T-cell development in the neonatal and juvenile thymus

doi: 10.1038/s41598-017-00349-8

Figure Lengend Snippet: FGF21 treatment alters T-cell development in FTOC. WT and Fgf21 KO foetal thymi were cultured with or without recombinant human FGF21 protein (500 ng/ml) for 14 days. ( A ) Total cell numbers of thymocytes in FTOCs are presented. Flow cytometry analysis of CD4/8 ( B ) and TCRβ/CD69 ( C ) distribution and thymocyte subset number ( D and E ). ( F ) Representative flow cytometry data for each FTOC. ( G ) Numbers of mTEC (CD45 − EpCAM + UEA-1 + ), cTEC (CD45 − EpCAM + UEA-1 − ), and non-TEC stromal cells (CD45 − EpCAM − ) are shown. Data represents the mean ± SD of two separate experiments. *,# P < 0.05, **,## P < 0.01, and *** P < 0.001 versus WT without rhFGF21, $ P < 0.05, and $$ P < 0.01 versus Fgf21 KO without rhFGF21.

Article Snippet: Medium containing 500 ng/ml of recombinant human FGF21 (R&D Systems) was replaced at 3-day intervals.

Techniques: Cell Culture, Recombinant, Flow Cytometry

Journal: Scientific Reports

Article Title: Fgf21 regulates T-cell development in the neonatal and juvenile thymus

doi: 10.1038/s41598-017-00349-8

Figure Lengend Snippet: FGF21 treatment results in increased apoptosis of immature thymocytes. FTOC was performed in the presence or absence of recombinant human FGF21 (500 ng/ml) for 14 days. Annexin V staining was performed to detect apoptotic T cells by flow cytometry. ( A ) Representative histograms show the percentage of Annexin V + cells in gated thymocyte subsets. ( B ) Graph showing the analysis of the percentage of Annexin V + cells in thymocytes from FTOCs. All data shown are the mean ± SD. **P < 0.01 versus control.

Article Snippet: Medium containing 500 ng/ml of recombinant human FGF21 (R&D Systems) was replaced at 3-day intervals.

Techniques: Recombinant, Staining, Flow Cytometry, Control

Journal: Scientific Reports

Article Title: Fgf21 regulates T-cell development in the neonatal and juvenile thymus

doi: 10.1038/s41598-017-00349-8

Figure Lengend Snippet: Fgf21 induced by protein-free diet does not affect thymic change by protein malnutrition. ( A ) Serum Fgf21 levels of 4-week-old C57BL/6 mice fed for 1 week with normal chow diet (NC) or protein-free diet (PF) were determined with ELISA assays. Data shown are the mean ± SD from ≧6 mice. ** P < 0.001 versus NC. ( B ) Relative expression levels of Fgf21 mRNA in the liver, thymus, subcutaneous white adipose tissue (sWAT), and skeletal muscle from mice fed with NC or PF. Data shown are the mean ± SD from ≧6 mice. ** P < 0.01, ** P < 0.001 versus NC. ( C–F ) Body weight ( C ), blood glucose ( D ), thymic weight ( E ), and thymic cell number ( F ) of WT and Fgf21 KO mice fed with NC or PF. Data shown are the mean ± SD from ≧5 mice. * P < 0.05, # P < 0.05 versus WT and Fgf21 KO mice fed with NC, respectively. $ P < 0.05 versus WT mice fed with PF. ( G and H ) Thymocytes from WT and Fgf21 KO mice fed with NC or PF were stained with anti-CD4 and anti-CD8 mAb ( G ), or anti-TCRβ and anti-CD69 mAb ( H ). All data shown are the mean ± SD from ≧5 mice. * P < 0.05, # P < 0.05 versus WT and Fgf21 KO mice fed with NC, respectively. $ P < 0.05 versus WT mice fed with PF.

Article Snippet: Medium containing 500 ng/ml of recombinant human FGF21 (R&D Systems) was replaced at 3-day intervals.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Staining

Journal: Frontiers in Endocrinology

Article Title: Fibroblast Growth Factor 21-Null Mice Do Not Exhibit an Impaired Response to Fasting

doi: 10.3389/fendo.2016.00077

Figure Lengend Snippet: FGF21 is primarily expressed in the liver and is dispensable for the physiological response to fasting . WT and FGF21KO animals were either allowed ad libidum access to food (fed) or subjected to a 24-h fast (fasted). (A) Plasma FGF21 levels were measured in WT animals. (B) FGF21 mRNA and (C) protein expression were measured in liver and adipose tissue from WT animals. (D) Blood glucose was assessed via glucometer in each cohort. Plasma was collected from WT and FGF21KO animals and used to determine circulating levels of (E) insulin, (F) βHB, (G) triglycerides, and (H) free fatty acids. The concentration of (I) triglycerides and (J) glycogen was measured in the livers of WT and FGF21KO animals. (K) A pyruvate tolerance test was performed on a separate cohort of WT and FGF21KO animals by administration of 2 g/kg sodium pyruvate following a 22-h fast. * p < 0.05 vs. fed; α, p < 0.05 vs. WT Fed; β, p < 0.05 vs. FGF21KO fed; φ, p < 0.05 vs. WT fasted; #, p < 0.05 vs. WT. No statistical difference was observed in fed measurements between genotypes.

Article Snippet: Male C57Bl/6J and FGF21 knockout (FGF21KO) mice (Taconic Farms) were maintained on a standard chow diet consisting of 13% fat, 67% carbohydrate, and 20% protein caloric content (2014; Harlan Teklad, Madison, WI, USA) and had free access to food and water before randomization by weight.

Techniques: Clinical Proteomics, Expressing, Concentration Assay

Journal: Frontiers in Endocrinology

Article Title: Fibroblast Growth Factor 21-Null Mice Do Not Exhibit an Impaired Response to Fasting

doi: 10.3389/fendo.2016.00077

Figure Lengend Snippet: Oxidative and gluconeogenic gene expression is attenuated in the liver, while lipase gene expression and activity in WAT remains unaltered in fasted FGF21KO mice . (A) In the liver, expressions of genes associated with fatty acid oxidation (CPT1a, ACADS, ACADM, ACADL, ACADVL, HADH), fatty acid import (CD36), gluconeogenesis (G6PC, PCK1), and ketone body production (HMGCS2, BDH1) were measured. (B) Expressions of genes associated with lipolysis (HSL, ATGL, LPL), fatty acid synthesis (FASN), fatty acid import (CD36), and thermogenesis (CIDEA, UCP1) were measured. In adipose tissue: α, p < 0.05 vs. WT Fed; β, p < 0.05 vs. FGF21KO fed; φ, p < 0.05 vs. WT fasted. No statistical differences were observed between genotypes in fed expression of any gene measured. (C) The amount of total and phosphorylated HSL was assessed by Western blot in WAT of WT and FGF21KO animals in both the fed and fasted state.

Article Snippet: Male C57Bl/6J and FGF21 knockout (FGF21KO) mice (Taconic Farms) were maintained on a standard chow diet consisting of 13% fat, 67% carbohydrate, and 20% protein caloric content (2014; Harlan Teklad, Madison, WI, USA) and had free access to food and water before randomization by weight.

Techniques: Gene Expression, Activity Assay, Expressing, Western Blot

Journal: Frontiers in Endocrinology

Article Title: Fibroblast Growth Factor 21-Null Mice Do Not Exhibit an Impaired Response to Fasting

doi: 10.3389/fendo.2016.00077

Figure Lengend Snippet: Endogenous FGF21 is not required for proper lipid and carbohydrate metabolism during the fed to fasted transition . At the beginning of the light cycle, WT animals had their food removed and were administered 5 mg/kg ΔN17 once every 3 h for 12 h. (A) Blood glucose measurements were taken via glucometer at the specified time points. Circulating levels of (B) insulin, (C) βHB, (D) triglycerides, and (E) free fatty acids were determined from plasma samples collected at the end of the study. Liver samples were taken at the end of the study and used to determine the concentrations of (F) triglycerides and (G) glycogen. (H) In the liver, expression of FGF21 as well as genes associated with fatty acid oxidation (CPT1a, ACADS, ACADM, ACADL, ACADVL, HADH), fatty acid import (CD36), gluconeogenesis (G6PC, PCK1), and ketone body production (HMGCS2, BDH1) were measured. (I) Expression of FGF21 in addition to genes associated with lipolysis (HSL, ATGL, LPL), fatty acid synthesis (FASN), fatty acid import (CD36), and thermogenesis (CIDEA, UCP1) were measured in adipose tissue. * p < 0.05 vs. PBS.

Article Snippet: Male C57Bl/6J and FGF21 knockout (FGF21KO) mice (Taconic Farms) were maintained on a standard chow diet consisting of 13% fat, 67% carbohydrate, and 20% protein caloric content (2014; Harlan Teklad, Madison, WI, USA) and had free access to food and water before randomization by weight.

Techniques: Clinical Proteomics, Expressing

Journal: Frontiers in Endocrinology

Article Title: Fibroblast Growth Factor 21-Null Mice Do Not Exhibit an Impaired Response to Fasting

doi: 10.3389/fendo.2016.00077

Figure Lengend Snippet: Blockade of FGF21 signaling during late fasting does not impair the fasted response . WT mice were fasted for 21 h, administered 5 mg/kg ΔN17, and then subjected to a PTT 1 h later by administration of 2 g/kg sodium pyruvate. (A) Blood glucose was measured at the specified time points via glucometer. A separate cohort of WT mice was fasted for 23 h, administered 5 mg/kg ΔN17, and then sacrificed 1 h later. (B) Blood glucose was assessed via glucometer. Plasma samples were collected and used to determine circulating levels of (C) insulin, (D) βHB, (E) triglycerides, and (F) free fatty acids. Hepatic (G) Triglyceride and (H) glycogen content was measured. (I) In the liver, expression of FGF21 as well as genes associated with fatty acid oxidation (CPT1a, ACADS, ACADM, ACADL, ACADVL, HADH), fatty acid import (CD36), gluconeogenesis (G6PC, PCK1), and ketone body production (HMGCS2, BDH1) were measured. (J) Expression of FGF21 in addition to genes associated with lipolysis (HSL, ATGL, LPL), fatty acid synthesis (FASN), fatty acid import (CD36), and thermogenesis (CIDEA, UCP1) were measured in adipose tissue. * p < 0.05 vs. PBS.

Article Snippet: Male C57Bl/6J and FGF21 knockout (FGF21KO) mice (Taconic Farms) were maintained on a standard chow diet consisting of 13% fat, 67% carbohydrate, and 20% protein caloric content (2014; Harlan Teklad, Madison, WI, USA) and had free access to food and water before randomization by weight.

Techniques: Clinical Proteomics, Expressing

Journal: Stroke and vascular neurology

Article Title: Plasma from healthy donors protects blood-brain barrier integrity via FGF21 and improves the recovery in a mouse model of cerebral ischaemia.

doi: 10.1136/svn-2020-000774

Figure Lengend Snippet: Figure 4 Healthy plasma increased FGF21 and its' receptors expression in the tMCAO mouse brain photomicrograph showed that Western blot images of FGF21 bands (A) and bar graph (B) showed semiquantification of FGF21 protein expression in sham, saline-injected and plasma-injected mouse brain at 3 days of tMCAO. Data are mean±SD, n=3 per group. *p<0.05, plasma-injected versus control mice. RT-PCR quantification of FGF21 (C), β-klotho (D), FGFR1 (E), FGFR2 (F) and FGFR3 (G) mRNA expression in the sham, saline-injected and plasma-injected mouse brain at 3 days of tMCAO. Data are mean±SD, n=4 per group. *p<0.05; ** p<0.01, plasma-injected versus control. FGF21, fibroblast growth factor 21; FGFR1, fibroblast growth factor receptor; tMCAO, transient middle cerebral artery occlusion.

Article Snippet: FGF21- KO mice (C57BL/6 background) purchased from Cyagen Biosciences Inc. Suzhou, China.

Techniques: Clinical Proteomics, Expressing, Western Blot, Saline, Injection, Control, Reverse Transcription Polymerase Chain Reaction

Journal: Stroke and vascular neurology

Article Title: Plasma from healthy donors protects blood-brain barrier integrity via FGF21 and improves the recovery in a mouse model of cerebral ischaemia.

doi: 10.1136/svn-2020-000774

Figure Lengend Snippet: Figure 5 Depletion of FGF21 in plasma reversed the beneficial effect of healthy plasma in the tMCAO mice bar graphs showed that neurological severity score (A), rotarod test (B), EBST (C) and corner test (D) in wild-type mice that treated with plasma from wild-type mice (WT+WT); wild-type mice that treated with plasma from FGF21-/- mice (WT+ FGF21-/- plasma); and FGF21- /- mice that treated with wild-type plasma (FGF21-/-+ WT plasma) at 1, 3, 7 and 14 days after tMCAO. (E, F) Images of cresyl violet-stained brain sections of WT+ saline group, WT+WT group, WT+ FGF21-/- plasma group, and FGF21-/-+ WT plasma group at 3 and 14 days of tMCAO. Dashed line showed infarct area in the ipsilateral hemisphere of the brain. Semiquantification of brain oedema and infarct volume in WT+ saline group, WT+WT group, WT+ FGF21-/- plasma group, and FGF21-/-+ WT plasma group at 3 days of tMCAO (G, H). Bbar graph showed that the brain atrophy volume and the volume ratio of the ipsilateral hemisphere and contralateral hemisphere in the WT+ saline group, WT+WT group, WT+ FGF21-/- plasma group and FGF21-/-+ WT plasma group at 14 days in the tMCAO mouse brain (I, J). Data are mean±SD, n=10 per group. *p<0.05, **p<0.01, ***p<0.001, FGF21-treated versus control mice. EBST, elevated body swing test; FGF21, fibroblast growth factor 21; tMCAO, transient middle cerebral artery occlusion.

Article Snippet: FGF21- KO mice (C57BL/6 background) purchased from Cyagen Biosciences Inc. Suzhou, China.

Techniques: Clinical Proteomics, Staining, Saline, Control

Journal: Stroke and vascular neurology

Article Title: Plasma from healthy donors protects blood-brain barrier integrity via FGF21 and improves the recovery in a mouse model of cerebral ischaemia.

doi: 10.1136/svn-2020-000774

Figure Lengend Snippet: Figure 6 Depletion of FGF21 in plasma diminished the beneficial effects of plasma-induced BBB protection in mice after 3 days of tMCAO coimmunostaining of endothelial cell marker CD31(red) with BBB (green) ZO-1(A), occludin (C) and claudin-5 (E) in the peri-infarct area of the striatum in wild-type mice that treated with saline (WT+ saline), wild-type mice that treated with wild mouse plasma (WT+WT), wild-type mice that treated with plasma from FGF21-/- mice (WT+ FGF21-/- plasma) and FGF21- /- KO mice that treated with wild-type plasma (FGF21-/- + WT plasma) at 3 days of tMCAO. Arrows indicate the gaps in BBB tight junction proteins, scale bar=10 μm. Quantification of gaps that formed on BBB tight junction proteins ZO-1(B), occludin (D) and claudin-5 (F) in the WT+ saline group, WT+WT group, WT+ FGF21-/- plasma group, and FGF21-/-+ WT plasma group at 3 days of tMCAO. Data are mean±SD, n=3 per group. **p<0.01. ***p<0.001. (G) Photomicrographs of IgG in the perifocal area of WT+ saline group, WT+WT group, WT+ FGF21-/- plasma group and FGF21-/-+ WT plasma group at 3 days after stroke. Scale bar=75 μm. (H) Bar graph showed semiquantification of IgG extravasation in the WT+ Saline group, WT+WT group, WT+FGF21-/- plasma group and FGF21-/-+WT plasma group. Data are presented as mean±SD, n=5 per group. * p<0.05; ****p<0.001, plasma versus control. BBB, blood-brain barrier; FGF21, fibroblast growth factor 21; tMCAO, transient middle cerebral artery occlusion; ZO-1, zonula occludens-1.

Article Snippet: FGF21- KO mice (C57BL/6 background) purchased from Cyagen Biosciences Inc. Suzhou, China.

Techniques: Clinical Proteomics, Marker, Saline, Control