recombinant fgf21 protein  (R&D Systems)

Journal: Journal of Inflammation Research

Article Title: FGF21 Exacerbates Obesity-Induced Airway Hyperresponsiveness and FGFR1-Dependent Mast Cell Activation in Mice

doi: 10.2147/JIR.S570000

Figure Lengend Snippet: FGF21 is upregulated in serum and BALF of obese mice, and positively correlated with airway resistance. Mice were fed either a standard chow diet (lean) or a high-fat diet (obese) for 16 weeks prior to analysis. ( A ) Measurements of body weight from the lean and obese group mice. ( B ) Representative pictures of lean and obese mice. ( C ) Levels of serum FGF21 in lean and obese mice were measured by ELISA (n=10). ( D ) Levels of BALF FGF21 in lean and obese mice were measured by ELISA (n=10). ( E ) DIO mice exhibit pronounced AHR. Results showed the changes in specific airway resistance (sRaw) as a measure of AHR. ( F ) Pearson’s correlation tests results represent FGF21 level in serum of obese mice was positively correlated with sRaw (C methacholine :50.0 mg/mL). ( G ) Pearson’s correlation tests results represent FGF21 level in BALF of obese mice was positively correlated with sRaw (C methacholine :50.0 mg/mL). Data are mean ± SEM, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant mouse FGF21 was purchased from MedChemExpress (HY- P72651 ).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: FGF21 Exacerbates Obesity-Induced Airway Hyperresponsiveness and FGFR1-Dependent Mast Cell Activation in Mice

doi: 10.2147/JIR.S570000

Figure Lengend Snippet: FGF21 level is increased in serum from obese patients with asthma and positively correlated with reduced pulmonary function. ( A ) Serum FGF21 levels in lean patients with asthma and obese patients with asthma were measured by ELISA (n=20-23). Data are mean ± SEM, * p < 0.05, *** p < 0.001 ( B ) Multivariable linear regression results represent serum FGF21 level was negatively correlated with FEV1%. ( C ) Multivariable linear regression results represent serum FGF21 level was negatively correlated with FEV1/FVC%, n=43.

Article Snippet: Recombinant mouse FGF21 was purchased from MedChemExpress (HY- P72651 ).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Inflammation Research

Article Title: FGF21 Exacerbates Obesity-Induced Airway Hyperresponsiveness and FGFR1-Dependent Mast Cell Activation in Mice

doi: 10.2147/JIR.S570000

Figure Lengend Snippet: Recombinant FGF21 aggravates AHR in obese mice, while Anti-FGF21 ameliorates obesity-induced AHR and inhibits mast cell infiltration. ( A ) The schematic diagram for recombinant FGF21 treatment in DIO mice. ( B ) Changes of airway resistance in DIO mice treated with recombinant FGF21. ( C ) Representative immunohistochemical images (× 400) for lung tissue sections staining of chymase. ( D ) Quantitative analysis of chymase staining. ( E ) The schematic diagram for Anti-FGF21 treatment in lean mice or DIO mice. ( F ) Changes of lung resistance (R L ) in DIO mice treated with the Anti-FGF21. ( G ) Representative immunohistochemical images (× 400) for lung tissue sections staining of chymase. ( H ) Quantitative analysis of chymase staining. n=5. Data are mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001. IgG: the normal IgG control antibody.

Article Snippet: Recombinant mouse FGF21 was purchased from MedChemExpress (HY- P72651 ).

Techniques: Recombinant, Immunohistochemical staining, Staining, Control

Journal: Journal of Inflammation Research

Article Title: FGF21 Exacerbates Obesity-Induced Airway Hyperresponsiveness and FGFR1-Dependent Mast Cell Activation in Mice

doi: 10.2147/JIR.S570000

Figure Lengend Snippet: FGF21 facilitates mast cell activation through up-regulating cholesterol biosynthesis. ( A ) Release of β-hexosaminidase from LAD2 and P815 cells pre-activated with compound 48/80 following 24 h treatment with recombinant FGF21 (100–400 ng/mL). ( B ) Release of histamine from LAD2 and P815 cells pre-activated with compound 48/80 following 24 h treatment with recombinant FGF21 (100–400 ng/mL). ( C ) Measurement of cellular calcium concentration in mast cells pre-activated with compound 48/80 using fluorescent probe Fluo-4 AM following 24 h treatment with recombinant FGF21 (200 ng/mL). ( D ) Quantitative analysis of fluorescence intensity of Fluo-4. ( E ) Filipin III staining of cholesterol in mast cells pre-activated with compound 48/80 following 24 h treatment with recombinant FGF21 (200 ng/mL). ( F ) Quantitative analysis of fluorescence intensity of Filipin III. ( G ) Expression levels of cholesterol biosynthesis genes in LAD2 cells. ( H ) Expression level of SREBF1 . ( I ) Quantitative analysis of fluorescence intensity of Filipin III in siRNA pretreated cells. ( J ) Representative images of Filipin III staining in siRNA-NC or siRNA- SREBF1 pretreated cells. ( K ) Release rate of β-hexosaminidase from LAD2 cells treated with siRNA and FGF21. ( L ) Measurement of cellular calcium concentration in LAD2 cells using fluorescent probe Fluo-4 AM. ( M ) Quantitative analysis of fluorescence intensity of Fluo-4 in LAD2 cells treated with siRNA and FGF21. n=3. Data are mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001, ns, not significant.

Article Snippet: Recombinant mouse FGF21 was purchased from MedChemExpress (HY- P72651 ).

Techniques: Activation Assay, Recombinant, Concentration Assay, Fluorescence, Staining, Expressing

Journal: Journal of Inflammation Research

Article Title: FGF21 Exacerbates Obesity-Induced Airway Hyperresponsiveness and FGFR1-Dependent Mast Cell Activation in Mice

doi: 10.2147/JIR.S570000

Figure Lengend Snippet: FGF21 promoted cholesterol synthesis and mast cell activation in a FGFR1-dependent manner. ( A ) Expression of FGFR1, FGFR2 and FGFR3 in lung tissues. (Data from Human Protein Atlas, http://www.proteinatlas.org/ ). ( B ) The mRNA expression of FGFR1, FGFR2 , and FGFR3 in LAD2 cells. Relative expression levels were normalized to GAPDH . ( C ) Release rate of β-hexosaminidase from LAD2 cells treated with FGFR1 inhibitor PD173074 . ( D ) Measurement of cellular calcium concentration in PD173074 treated LAD2 cells using fluorescent probe Fluo-4 AM. ( E ) Quantitative analysis of fluorescence intensity of Fluo-4 in LAD2 cells. ( F ) Expression levels of cholesterol biosynthesis genes in PD173074 treated LAD2 cells. ( G ) Representative images of Filipin III staining in PD173074 treated LAD2 cells. ( H ) Quantitative analysis of fluorescence intensity of Filipin III in PD173074 treated cells. n=3. Data are mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant mouse FGF21 was purchased from MedChemExpress (HY- P72651 ).

Techniques: Activation Assay, Expressing, Concentration Assay, Fluorescence, Staining

Journal: Journal of Inflammation Research

Article Title: FGF21 Exacerbates Obesity-Induced Airway Hyperresponsiveness and FGFR1-Dependent Mast Cell Activation in Mice

doi: 10.2147/JIR.S570000

Figure Lengend Snippet: Schematic of the role of FGF21 in obesity induced AHR. FGF21 promotes AHR in obese mice through increasing cholesterol synthesis and facilitating mast cell activation in a FGFR1-dependent manner.

Article Snippet: Recombinant mouse FGF21 was purchased from MedChemExpress (HY- P72651 ).

Techniques: Activation Assay

Journal: Physiological Reports

Article Title: Identification of FGF21 ‐inducing rare sugars that reduces sugar appetite in male BL /6 mice

doi: 10.14814/phy2.70618

Figure Lengend Snippet: Identification of FGF21‐inducing rare sugars and their effects on blood glucose. (a–d) FGF21 level of mouse primary hepatocytes treated for 24 h with vehicle (distilled water‐negative control) or 25 mM D‐glucose, D‐fructose (positive control), or rare sugars (46 samples). n = 3 mice. (e) Plasma FGF21 levels over 24 h after gastric gavage of vehicle (distilled water), D‐glucose, D‐tagatose, D‐allulose, or D‐sorbitol at 5 g/kg in nine‐week‐old male WT mice with ad libitum access to normal chow (NC) diet. n = 4 mice per group. (f) Area under the curve (AUC) over 24 h of data in (e). n = 4 mice per group. (g) Blood glucose levels of the mice receiving the same dose in (e) of either vehicle (distilled water), D‐glucose, D‐tagatose, D‐allulose, or D‐sorbitol after 16 h fasting. n = 4 mice per group. Statistical analyses were done by one‐way ANOVA for (a–d, f) and repeated measures ANOVA for (e, g), followed by Tukey's HSD test. ** p < 0.01; **** p < 0.0001.

Article Snippet: Mouse FGF21 protein levels in each plasma sample were determined using a Mouse/Rat FGF‐21 Quantikine ELISA Kit (R&D Systems) according to the manufacturer's instructions.

Techniques: Negative Control, Positive Control, Clinical Proteomics

Journal: Physiological Reports

Article Title: Identification of FGF21 ‐inducing rare sugars that reduces sugar appetite in male BL /6 mice

doi: 10.14814/phy2.70618

Figure Lengend Snippet: FGF21‐inducing rare sugars activated oxytocin neurons of paraventricular nucleus of hypothalamus in BL/6 mice. (a–e) Immunostaining of mice brain after gastric gavage with either (a) vehicle (distilled water) or 5 g/kg body weight of D‐glucose (b), D‐tagatose (c), D‐allulose (d), or D‐sorbitol (e). Blue arrow ( ) indicates c‐Fos‐positive oxytocin neurons, shown by dark brown spot on the light brown neurons ( ). Magenta arrow ( ) indicates c‐Fos‐negative oxytocin neurons, shown by light brown anti‐oxytocin staining ( ). n = 4 mice per group. Scale bar represents 100 μ m. (f) Percentage of activated oxytocin neurons (c‐Fos positive oxytocin neurons) from the total of PVH oxytocin neurons in the mice receiving either vehicle (distilled water), D‐glucose, or FGF21‐inducing rare sugars in (a–e). Data are presented as box‐whiskers in (f), where the middle line represents median, bottom/ top edges represent 25th/75th percentile of the data, and whiskers represent maximum and minimum value of the data. Statistical analyses were done by one‐way ANOVA, followed by Dunnett's test.

Article Snippet: Mouse FGF21 protein levels in each plasma sample were determined using a Mouse/Rat FGF‐21 Quantikine ELISA Kit (R&D Systems) according to the manufacturer's instructions.

Techniques: Immunostaining, Staining

Journal: Physiological Reports

Article Title: Identification of FGF21 ‐inducing rare sugars that reduces sugar appetite in male BL /6 mice

doi: 10.14814/phy2.70618

Figure Lengend Snippet: Intragastric administration of FGF21‐inducing rare sugars reduced sucrose preference in BL/6 mice. (a–c) 50 mM sucrose intake in mice receiving 5 g/kg body weight of D‐allulose (a), D‐tagatose (b), or D‐sorbitol (c). (d–f) sucrose preference of the same mice in (a–c). n = 10 mice per group for (a, d) and n = 5 mice per group for (b, c, e, f). Data are presented as box‐whiskers, where the middle line represents the median, bottom/top edges represent the 25th/75th percentile of the data, and whiskers represent the maximum and minimum value of the data. Statistical analyses were done by Student's paired t ‐test between “pre” and “post” of each treatment.

Article Snippet: Mouse FGF21 protein levels in each plasma sample were determined using a Mouse/Rat FGF‐21 Quantikine ELISA Kit (R&D Systems) according to the manufacturer's instructions.

Techniques:

Journal: Physiological Reports

Article Title: Identification of FGF21 ‐inducing rare sugars that reduces sugar appetite in male BL /6 mice

doi: 10.14814/phy2.70618

Figure Lengend Snippet: Mixing FGF21‐inducing rare sugars into sucrose solution reduced solution intake and preference in BL/6 mice. (a–c) Intake of 100 mM (final concentration) sucrose solution that was mixed with either vehicle (distilled water), 600 mM (final concentration) of D‐glucose, or 600 mM (final concentration) of D‐allulose (a), D‐tagatose (b), or D‐sorbitol (c). (d–f) Sucrose preference of the same mice in (a–c). n = 6 mice per group. Data are presented as box‐whiskers, where the middle line represents the median, bottom/top edges represent the 25th/75th percentile of the data, and whiskers represent the maximum and minimum value of the data. Statistical analyses were done by repeated measures ANOVA, followed by Tukey's HSD test.

Article Snippet: Mouse FGF21 protein levels in each plasma sample were determined using a Mouse/Rat FGF‐21 Quantikine ELISA Kit (R&D Systems) according to the manufacturer's instructions.

Techniques: Concentration Assay