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: Science Advances

Article Title: Sustained release of a GLP-1 and FGF21 dual agonist from an injectable depot protects mice from obesity and hyperglycemia

doi: 10.1126/sciadv.aaz9890

Figure Lengend Snippet: Six-week-old db/db mice ( n = 6 to 7) were subcutaneously injected with ELP-FGF21 (1000 nmol/kg), GLP1-ELP (1000 nmol/kg), or GLP1-ELP and ELP-FGF21 (1000 nmol/kg each). Ambient blood glucose levels ( A ) and body weights ( B ) were measured 48 hours after injection and reported as a magnitude change from pretreatment baseline and a percentage change from preinjection weight. Data are presented as means ± SEM and were analyzed by one-way ANOVA, followed by Tukey’s tests. *, treatment compared to vehicle; ^, comparisons between treatments; ^ P < 005, ** P < 0.01, *** P < 0.001, and ****/^^^^ P < 0.0001. NS, not significant ( P > 0.05).

Article Snippet: After serum starvation for 6 hours, cells were treated with serial dilutions of FGF21-containing fusion proteins or native mouse FGF21 (ProSpec-Tany, East Brunswick, NJ) for 5 min.

Techniques: Injection

Journal: Science Advances

Article Title: Sustained release of a GLP-1 and FGF21 dual agonist from an injectable depot protects mice from obesity and hyperglycemia

doi: 10.1126/sciadv.aaz9890

Figure Lengend Snippet: ( A ) SDS-PAGE analysis of the 72-kDa fusion protein following recombinant expression in E. coli and ITC-based purification. 1, molecular weight ladder (kilodaltons); 2, cell lysate; 3, insoluble lysate fraction; 4, soluble lysate fraction; 5, hot spin supernatant; 6 to 8, ITC rounds 1 to 3. ( B and C ) In vitro GLP-1 and FGF21 activity assays for GLP1-ELP-FGF21. GLP-1R agonism (B) was measured by quantifying adenosine 3′,5′-monophosphate (cAMP) production in human embryonic kidney (HEK) 293 cells stably expressing the GLP-1R and the cAMP-inducible luciferase reporter. Cells were stimulated for 5 hours with GLP1-ELP-FGF21, the GLP1-ELP single-agonist control, or native GLP-1. FGF21 receptor agonism (C) was measured by quantifying extracellular signal–regulated kinase 1/2 (ERK1/2) phosphorylation in HEK293 cells stably expressing FGF receptor 1 (FGFR1) and β-klotho and by normalizing phospho-ERK1/2 to total ERK1/2. Cells were stimulated for 5 min with GLP1-ELP-FGF21, the ELP-FGF21 single-agonist control, or native FGF21. Data are presented as means ± SEM, n = 3. ( D to F ) LCST phase transition behavior of GLP1-ELP-FGF21. (D) The optical density (OD) at 350 nm of GLP1-ELP-FGF21 at the indicated concentration in phosphate-buffered saline (PBS) was measured as a function of temperature, with temperature ramped at a rate of 1°C/min. (E) Turbidity versus temperature scans were repeated as in (D) for the indicated concentrations ( n = 3). T t values were defined as the temperature corresponding to the 50% maximum OD and are plotted as a function of fusion protein concentration. The horizontal dashed line indicates the approximate temperature of the subcutaneous space in a mouse . (F) A turbidity scan was repeated for GLP1-ELP-FGF21 at an injection-relevant concentration (150 μM), with the temperature ramped up to 37°C and then down to 20°C.

Article Snippet: After serum starvation for 6 hours, cells were treated with serial dilutions of FGF21-containing fusion proteins or native mouse FGF21 (ProSpec-Tany, East Brunswick, NJ) for 5 min.

Techniques: SDS Page, Recombinant, Expressing, Purification, Molecular Weight, In Vitro, Activity Assay, Stable Transfection, Luciferase, Control, Phospho-proteomics, Sublimation, Concentration Assay, Saline, Protein Concentration, Injection

Journal: Science Advances

Article Title: Sustained release of a GLP-1 and FGF21 dual agonist from an injectable depot protects mice from obesity and hyperglycemia

doi: 10.1126/sciadv.aaz9890

Figure Lengend Snippet: ( A to D ) Six-week-old db/db mice ( n = 3 to 4) received a single subcutaneous injection of GLP1-ELP-FGF21 at the indicated dose or vehicle. Ambient blood glucose levels were measured every 24 hours until animals returned to baseline levels and are reported as blood glucose versus time AUC or raw values (A and B). Body weights were recorded daily and are reported as a percentage change from preinjection weight over time (C) or on day 7 after injection (D). ( E ) GLP1-ELP-FGF21 (1000 nmol/kg) was subcutaneously administered to 6-week-old db/db mice ( n = 4) as a radiolabeled protein, blood samples were collected at indicated time points following injection, and plasma gamma counts were correlated to fusion protein concentration. Regression curves were fit to the terminal portion of the dataset, and data could be described by either a first-order (dotted) or a zero-order (solid) elimination model. Data are presented as means ± SEM and were analyzed by one-way ANOVA, followed by Dunnett’s tests. * P < 0.05 and ** P < 0.01.

Article Snippet: After serum starvation for 6 hours, cells were treated with serial dilutions of FGF21-containing fusion proteins or native mouse FGF21 (ProSpec-Tany, East Brunswick, NJ) for 5 min.

Techniques: Injection, Clinical Proteomics, Protein Concentration

Journal: Science Advances

Article Title: Sustained release of a GLP-1 and FGF21 dual agonist from an injectable depot protects mice from obesity and hyperglycemia

doi: 10.1126/sciadv.aaz9890

Figure Lengend Snippet: ( A ) A mixed ELP depot releases GLP-1 and FGF21 at different rates. Six-week-old db/db mice ( n = 4 to 5) received a single subcutaneous injection consisting of a 1:1 mixture of the synthesized GLP1-ELP and previously reported ELP-FGF21 . The mixture was tested once when GLP1-ELP was radiolabeled and once when ELP-FGF21 was radiolabeled. Fusions were injected at 200 μM and dosed at 1000 nmol/kg each of GLP1-ELP and ELP-FGF21. Blood samples were collected at indicated time points following injection, and plasma gamma counts were measured and correlated to fusion protein concentration. Lines represent regression curves fit to the terminal portion of each dataset. Data can be described by both the first-order (dotted) and zero-order (solid) elimination models. ( B to G ) Six-week-old db/db mice ( n = 6 to 7) were treated weekly for 4 weeks with GLP1-ELP-FGF21, GLP1-ELP, ELP-FGF21, or PBS vehicle. Fusion proteins were administered subcutaneously at 1000 nmol/kg. (B to D) Glucose challenge. Seventy-two hours after the first treatment cycle, mice were fasted 5 hours, baseline blood glucose levels were measured (B), and mice were injected intraperitoneally with glucose (0.75 g/kg). Blood glucose levels were measured at indicated time points (C), and blood glucose versus time AUC values were calculated (D). (E and F) Body weights and food consumption were measured every 1 to 2 days and are reported as percentage change from preinjection weights (E) and cumulative food intake per mouse (F). (G) Before the first treatment (day 0) and 6 days following the final treatment (day 27), percent glycated hemoglobin A1c (%HbA1c) was measured in all cohorts and reported as a magnitude change from prestudy values. Data are presented as means ± SEM and were analyzed by one-way ANOVA or two-way repeated-measures ANOVA, followed by Dunnett’s tests. * P < 0.05, *** P < 0.001, and **** P < 0.0001.

Article Snippet: After serum starvation for 6 hours, cells were treated with serial dilutions of FGF21-containing fusion proteins or native mouse FGF21 (ProSpec-Tany, East Brunswick, NJ) for 5 min.

Techniques: Injection, Synthesized, Clinical Proteomics, Protein Concentration

Journal: Science Advances

Article Title: Sustained release of a GLP-1 and FGF21 dual agonist from an injectable depot protects mice from obesity and hyperglycemia

doi: 10.1126/sciadv.aaz9890

Figure Lengend Snippet: ( A to F ) Six-week-old db/db mice ( n = 7) received a single subcutaneous injection of GLP1-ELP-FGF21 (1000 nmol/kg), a 1:1 equimolar mixture of GLP1-ELP and ELP-FGF21, or an ELP-only control. Ambient blood glucose levels were measured every 24 hours until animals returned to baseline levels and are reported as raw values (A) or blood glucose versus time AUC (B). (C to E) Glucose challenge. Six days after treatment, mice were fasted 5 hours, baseline blood glucose levels were measured (C), and mice were injected intraperitoneally with glucose (0.75 g/kg). Blood glucose levels were measured at indicated time points (D), and blood glucose versus time incremental AUC (iAUC) values were calculated (E). Body weights were recorded daily and are reported as a percentage change from preinjection weight over time (F). ( G ) Six-week-old db/db mice ( n = 6 to 7) received a single subcutaneous injection of GLP1-ELP-FGF21 (1000 nmol/kg), a 1:1 equimolar mixture of GLP1-ELP and ELP-FGF21, or vehicle control. Food consumption was measured every 1 to 2 days and is reported as cumulative food intake per mouse. Data are presented as means ± SEM and were analyzed by one-way ANOVA or two-way repeated-measures ANOVA, followed by Dunnett’s tests. * P < 0.05 and **** P < 0.0001.

Article Snippet: After serum starvation for 6 hours, cells were treated with serial dilutions of FGF21-containing fusion proteins or native mouse FGF21 (ProSpec-Tany, East Brunswick, NJ) for 5 min.

Techniques: Injection, 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 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: Molecular metabolism

Article Title: Fibroblast growth factor 21 has no direct role in regulating fertility in female mice.

doi: 10.1016/j.molmet.2016.05.010

Figure Lengend Snippet: Figure 3: High fat diet reverses the infertility in FGF21-Tg mice. A) Body weight of WT and FGF21-Tg mice fed either with chow or HFD (n ¼ 4e6). B) Body composition as represented as absolute fat and lean mass (n ¼ 6e7). C) Assessment of estrus cycle in FGF21-Tg mice fed either chow or HFD. (1: leukocytes [diestrus/metestrus], 2: cornified cells [estrus], 3: nucleated cells [proestrus]) D) Representative vaginal cytology from FGF21 Tg mice fed with HFD as (i) diestrus, (ii) proestrus, (iii) estrus, (iv) metestrus. Scale: 200 mm (ieiv). (n ¼ 8). E) Ovarian histology from FGF21-Tg mice; (i) chow, (ii) HFD, (iiieiv) HFD fed pregnant females. Corpus lutea (CL) are encircled with dotted lines. Scale: 1000 mm. Statistical significance was evaluated by Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Mouse FGF21 and human FGF21 proteins were measured by ELISA (R & D Systems, MN).

Techniques:

Journal: Molecular metabolism

Article Title: Fibroblast growth factor 21 has no direct role in regulating fertility in female mice.

doi: 10.1016/j.molmet.2016.05.010

Figure Lengend Snippet: Figure 5: Infertility of FGF21-Tg mice is independent of FGF21. Serum levels of total FGF21 (A) and active FGF21 (B) in FGF21-Tg mice fed with chow or HFD (n ¼ 9e10). AVP and Kiss-1 gene expression in SCN (C) and AVPV (D), respectively of WT and FGF21-Tg mice fed chow or HFD (n ¼ 8e10). All mice were in the diestrus phase. Statistical significance was evaluated by Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Mouse FGF21 and human FGF21 proteins were measured by ELISA (R & D Systems, MN).

Techniques: Gene Expression

Journal: Heliyon

Article Title: Homocysteine-induced endoplasmic reticulum stress activates FGF21 and is associated with browning and atrophy of white adipose tissue in Bhmt knockout mice

doi: 10.1016/j.heliyon.2023.e13216

Figure Lengend Snippet: FGF21 is increased in Bhmt- KO plasma and liver. (A) FGF21 levels are increased in plasma from Bhmt- KO mice when compared to Bhmt- WT. n = 8 per group. Results are represented as mean ± SEM. P values were calculated by t -test. (B) mRNA levels of Fgf21 in the liver are increased ∼2 fold. Relative quantitative values (normalized to 36B4) are reported as fold change. P values were calculated by t -test. N = 5 per group.

Article Snippet: For both plasma and liver, supernatant protein was quantified using BCA protein assay (Bio-Rad, Hercules, CA, USA) and diluted to equal concentrations before performing an enzyme-linked immunosorbent assay (ELISA) using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN) [ ].

Techniques: Clinical Proteomics

Journal: Heliyon

Article Title: Homocysteine-induced endoplasmic reticulum stress activates FGF21 and is associated with browning and atrophy of white adipose tissue in Bhmt knockout mice

doi: 10.1016/j.heliyon.2023.e13216

Figure Lengend Snippet: Schematic representation of the effects of the deletion of Bhmt in liver and iWAT compared to Bhmt- WT. The schema summarizes our new findings where deletion of Bhmt in mice increases homocysteine levels leading to endoplasmic reticulum (ER) stress. ER stress led to an increase in the cleaved CREBH protein levels, which acts as a transcription factor that binds the FGF21 promoter. FGF21 high levels exert their effects in iWAT.

Article Snippet: For both plasma and liver, supernatant protein was quantified using BCA protein assay (Bio-Rad, Hercules, CA, USA) and diluted to equal concentrations before performing an enzyme-linked immunosorbent assay (ELISA) using a Mouse/Rat FGF21 Quantikine ELISA kit (R&D Systems, Minneapolis, MN) [ ].

Techniques: