Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Antibodies utilized in the current study.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques:

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Short‐term semaglutide treatment improves glucose tolerance and reduces body weight in HFD‐challenged mice. (a) Diagram shows the animal experimental design. (b) Body weight changes during last 7 days in the three indicated groups. (c) Blood glucose level and area under the curve (AUC) during IPGTT. (d) Fasting (overnight) blood glucose levels at the end of the experiment for indicated groups. (e–g) Fasting plasma leptin (e), adiponectin (f), and FGF21 (g) levels. (h–m) Fat pad weights including epididymal (h, eWAT) and inguinal (i, iWAT) white adipose tissue and brown adipose tissue (j, BAT). (k) eWAT weight to body weight ratio. (l) iWAT weight to body weight ratio. (m) BAT weight to body weight ratio. Sema, semaglutide. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Clinical Proteomics

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment restores HFD‐induced attenuation on ERK phosphorylation to hFGF21 treatment in hepatocytes. (a) Western blotting show expression levels of indicated protein in MPH isolated from LFD‐fed mice after 1 h hFGF21 treatment with indicated dose. (b, c) Densitometric analyses for pERK (p44, Thr202) and pERK (p42, Tyr204) with indicated treatment. (d) Western blotting show expression levels of indicated protein in MPH isolated from HFD‐fed mice after 1 h hFGF21 treatment with indicated dose. (e, f) Densitometric analyses for pERK (p44, Thr202) and pERK (p42, Tyr204) with indicated treatment. (g) Western blotting show expression levels of indicated protein in MPH isolated from semaglutide‐treated mice after 1 h hFGF21 treatment with indicated dose. (h, i) Densitometric analyses for pERK (p44, Thr202) and pERK (p42, Tyr204) with indicated treatment. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Phospho-proteomics, Western Blot, Expressing, Isolation

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment restores the stimulatory effects of hFGF21 on its downstream target gene expressions in MPH. (a) qRT‐PCR show effect of indicated dose of hFGF21 treatment (4 h) on expression of cFos in MPH isolated from LFD‐fed, HFD‐fed, and semaglutide‐treated mice. (b) qRT‐PCR show effect of indicated dose of hFGF21 treatment (4 h) on expression of Egr1 in MPH isolated from LFD‐fed, HFD‐fed, and semaglutide‐treated mice. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Quantitative RT-PCR, Expressing, Isolation

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment improves FGF21 sensitivity and attenuates the effect of HFD feeding on hepatic gene expression. (a) Western blotting show expression levels of FGF21 in the liver of three indicated groups. (b–d) qRT‐PCR shows the comparison of expression levels on Fgf21 (b) and genes that encode its receptor, Fgfr1 (c) and co‐receptor Klb (d) in the liver of three indicated groups. (e) qRT‐PCR shows the comparison of expression levels of lipogenic and fatty acid oxidation genes in the liver of three indicated groups. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Gene Expression, Western Blot, Expressing, Quantitative RT-PCR, Comparison

Journal: Physiological Reports

Article Title: Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

doi: 10.14814/phy2.15620

Figure Lengend Snippet: Seven‐day semaglutide treatment recovers HFD‐induced alteration in a battery of adipose‐specific genes. (a–c) qRT‐PCR shows the comparison of expression levels on Fgfr1 (a), Klb (b) and Fgf21 (c) in the eWAT of three indicated groups. (d) qRT‐PCR shows the comparison of expression levels of a battery of adipose tissue‐specific genes in the eWAT of three indicated groups. (e) The diagram shows the observed effects of short term semaglutide treatment on mice fed with HFD. In MPH, seven‐day semaglutide treatment restores the response to hFGF21 treatment. In the liver and eWAT, the treatment improves FGF21 sensitivity and restores HFD‐induced attenuation on genes that involved in maintaining lipid homeostasis and other adipose tissue‐specific genes. Data are shown as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant human FGF21 (hFGF21) was purchased from Cayman Chemical.

Techniques: Battery, Quantitative RT-PCR, Comparison, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

doi: 10.3892/etm.2018.5840

Figure Lengend Snippet: Analysis of changes to hFGF-21 levels in PhbA. (A) Plasma concentration levels of hFGF-21 in PhbA and healthy volunteers. (B) Expression levels of hFGF-21 in liver cells isolated from clinical patients and healthy volunteers. **P<0.01. hFGF-21, human fibroblast growth factor-21; PhbA, patients with hepatitis B cirrhosis combined with adrenal insufficiency.

Article Snippet: Serum levels of hFGF-21 (cat. no. DF2100), tumor necrosis factor (TNF)-α (cat. no. DTA00C), interleukin (IL)-1β (cat. no. DLB50), IL-6 (cat. no. D6050) and IL-8 (cat. no. D8000C) were detected in PhbA and healthy volunteers using ELISA kits (IBL International GmbH, Hamburg, Germany), according to the manufacturer's protocol.

Techniques: Concentration Assay, Expressing, Isolation

Journal: Experimental and Therapeutic Medicine

Article Title: Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

doi: 10.3892/etm.2018.5840

Figure Lengend Snippet: Analysis of hFGF-21 expression levels in PPhbA and PhbA. (A) Plasma concentration levels of hFGF-21 between PPhbA and PhbA on day 30. (B) mRNA and protein expression levels of hFGF-21 in liver cells isolated from PPhbA and PhbA. **P<0.01. hFGF-21, human fibroblast growth factor-21; PPhbA, prognostic patients with hepatitis B cirrhosis combined with adrenal insufficiency; PhbA, patients with hepatitis B cirrhosis combined with adrenal insufficiency; PPhbA, patients who recovered form hepatitis B cirrhosis combined with adrenal insufficiency.

Article Snippet: Serum levels of hFGF-21 (cat. no. DF2100), tumor necrosis factor (TNF)-α (cat. no. DTA00C), interleukin (IL)-1β (cat. no. DLB50), IL-6 (cat. no. D6050) and IL-8 (cat. no. D8000C) were detected in PhbA and healthy volunteers using ELISA kits (IBL International GmbH, Hamburg, Germany), according to the manufacturer's protocol.

Techniques: Expressing, Concentration Assay, Isolation

Journal: Experimental and Therapeutic Medicine

Article Title: Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

doi: 10.3892/etm.2018.5840

Figure Lengend Snippet: Association of hFGF-21 plasma concentration with cellular immunity and humoral immunity in clinical patients. Relationship between concentration levels of hFGF-21 and (A) B lymphocyte level and (B) macrophagocyte level in patients during treatment. Association of hFGF-21 plasma concentration with percentage of (C) CD4 + and (D) CD8 + cells in serum in patients during treatment. hFGF-21, human fibroblast growth factor-21; CD, cluster of differentiation.

Article Snippet: Serum levels of hFGF-21 (cat. no. DF2100), tumor necrosis factor (TNF)-α (cat. no. DTA00C), interleukin (IL)-1β (cat. no. DLB50), IL-6 (cat. no. D6050) and IL-8 (cat. no. D8000C) were detected in PhbA and healthy volunteers using ELISA kits (IBL International GmbH, Hamburg, Germany), according to the manufacturer's protocol.

Techniques: Concentration Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

doi: 10.3892/etm.2018.5840

Figure Lengend Snippet: Effects of hFGF-21 on inflammatory cytokine expression levels in liver cells isolated form clinical patients. Plasma concentration levels of (A) TNF-α, (B) IL-6, (C) IL-1β and (D) IL-8 in PhbA and healthy volunteers. (E) mRNA and (F) protein expression levels of inflammatory cytokines in cells treated with hFGF-21 isolated from patients and the PPhbA control group. **P<0.01. hFGF-21, human fibroblast growth factor-21; PhbA, patients with hepatitis B cirrhosis combined with adrenal insufficiency; PPhbA, patients who recovered form hepatitis B cirrhosis combined with adrenal insufficiency; TNF, tumor necrosis factor; IL, interleukin.

Article Snippet: Serum levels of hFGF-21 (cat. no. DF2100), tumor necrosis factor (TNF)-α (cat. no. DTA00C), interleukin (IL)-1β (cat. no. DLB50), IL-6 (cat. no. D6050) and IL-8 (cat. no. D8000C) were detected in PhbA and healthy volunteers using ELISA kits (IBL International GmbH, Hamburg, Germany), according to the manufacturer's protocol.

Techniques: Expressing, Isolation, Concentration Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

doi: 10.3892/etm.2018.5840

Figure Lengend Snippet: Effects of hFGF-21 on inflammatory cytokine expression levels in renal epithelial cells isolated from clinical patients. (A) Gene expression levels of TNF-α, IL-6, IL-1β and IL-8 in renal epithelial cells isolated from clinical patients treated with hFGF-21 or not treated with hFGF-21. (B) Protein expression levels of TNF-α, IL-6, IL-1β and IL-8 in renal epithelial cells isolated from clinical patients treated with hFGF-21 or not treated with hFGF-21. **P<0.01. hFGF-21, human fibroblast growth factor-21; TNF, tumor necrosis factor; IL, interleukin.

Article Snippet: Serum levels of hFGF-21 (cat. no. DF2100), tumor necrosis factor (TNF)-α (cat. no. DTA00C), interleukin (IL)-1β (cat. no. DLB50), IL-6 (cat. no. D6050) and IL-8 (cat. no. D8000C) were detected in PhbA and healthy volunteers using ELISA kits (IBL International GmbH, Hamburg, Germany), according to the manufacturer's protocol.

Techniques: Expressing, Isolation

Journal: Experimental and Therapeutic Medicine

Article Title: Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

doi: 10.3892/etm.2018.5840

Figure Lengend Snippet: hFGF-21 regulates inflammatory cytokines through downregulation of the NF-κB-mediated TGF-β signaling pathway. (A) Effects of hFGF-21 on deposition of ECM and ROS expression levels in liver cells. (B) Effects of hFGF-21 on expression levels of TGF-β, NF-κB and KLF6 in liver cells. (C) Effects of hFGF-21 on expression levels of PDGF and EGF in liver cells. (D) Knockdown of NF-κB with Si-NF-κB increases TGF-β and KLF6 expression in liver cells. (E) Knockdown of NF-κB with Si-NF-κB suppresses PDGF and EGF expression in liver cells. (F) Effects of Si-NF-κB on protein expression levels of TNF-α, IL-6, IL-1β and IL-8 in liver cells. **P<0.01. hFGF-21, human fibroblast growth factor-21; NF, nuclear factor; TGF, transforming growth factor; ECM, extracellular matrix; ROS, reactive oxygen species; KLF6, Kruppel-like factor 6; PDGF, platelet-derived growth factor; EGF, epidermal growth factor; TNF, tumor necrosis factor; IL, interleukin; Si, small interfering RNA.

Article Snippet: Serum levels of hFGF-21 (cat. no. DF2100), tumor necrosis factor (TNF)-α (cat. no. DTA00C), interleukin (IL)-1β (cat. no. DLB50), IL-6 (cat. no. D6050) and IL-8 (cat. no. D8000C) were detected in PhbA and healthy volunteers using ELISA kits (IBL International GmbH, Hamburg, Germany), according to the manufacturer's protocol.

Techniques: Expressing, Derivative Assay, Small Interfering RNA

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: Construction of the hFGF21 expression vectors and schematic representation of the domain structure of fusion proteins. ( A ) The vector map of pHMGWA-hFGF21 for the Gateway® cloning method. Gene expression of the fusion proteins is under control of the IPTG-inducible T7 promoter. Ampicillin served as a selection marker. ( B ) Schematic structure of the eight tagged proteins His6-, Sumo-, Trx-, GST-, PDIb′a′-, MBP-, NusA-, and PDI-hFGF21 (total size). The arrow indicates the TEV protease cleavage site between the tags and hFGF21.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Expressing, Plasmid Preparation, Cloning, Gene Expression, Control, Selection, Marker

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: Expression levels of eight hFGF21 fusion proteins in E. coli BL21(DE3). Expression of the fusion proteins was induced by the addition of 0.5 mM IPTG at 37°C ( A ) or 18°C ( B ). The arrows indicate migration of hFGF21 fused with each tag in a polyacrylamide gel. M, molecular weight markers; C, total cell protein before IPTG induction as a negative control; I, whole cells after IPTG induction; P, the pellet fraction after cell sonication; S, the supernatant after cell sonication.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Expressing, Migration, Molecular Weight, Negative Control, Sonication

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: Expression and solubility levels of hFGF21 fused with eight different tags.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Expressing, Solubility

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: Purification of hFGF21 from soluble MBP-hFGF21 expressed in E. coli BL21(DE3). ( A ) The flowchart of the purification process. ( B ) The fusion protein and hFGF21 after tag cleavage were both purified by IMAC. M, molecular weight makers; lane 1, total cell protein before IPTG induction as a negative control; lane 2, whole-cell lysate after IPTG induction; lane 3, the soluble fraction after cell sonication; lane 4, the MBP-hFGF21 fusion protein purified by IMAC (64.4 kDa); lane 5, the MBP tag was cleaved by TEV protease: the MBP tag (40.3 kDa), hFGF21 (19.45 kDa); lane 6, the final hFGF21 product (19.45 kDa).

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Purification, Molecular Weight, Negative Control, Sonication

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: Prokaryotic purification of hFGF21 from solubly expressed MBP-FGF21.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Purification

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: SEC-HPLC analysis of hFGF21. The purified hFGF21 was analysed by HPLC using a protein-pak 300SW SEC 7.5 × 300 mm column to evaluate the purity. The x-axis indicates retention time (min), while the y-axis shows the absorbance at 280 nm (arbitrary units, AU). The main peak of pure hFGF21 appeared at 20.166 min.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Purification

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: Protein and disulphide bond identification using mass spectrometry-based analysis. ( A ) LC-MS/MS sequencing of peptides identifying the proteins was performed on samples digested with four enzymes (trypsin, Glu-C, ASP-N, and chymotrypsin). Percent coverage for protein identification is represented by the identified peptides in the total protein sequence; trypsin: 94.51%, Glu-C: 94.51%, Asp-N: 81.32%, chymotrypsin: 94.51%, and all four enzymes: 100%. Each sequence confidence is represented by PMSes as illustrated by high (green) and low (red) Percolator confidence scores. MALDI-TOF/TOF MS for purified hFGF21 in reducing condition ( B ) and non-reducing condition ( C ). The arrow indicates the peptide fragment with either the two IAA-alkylated cysteines or the formed disulphide bond.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Sequencing, Purification

Journal: Scientific Reports

Article Title: Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

doi: 10.1038/s41598-017-16167-x

Figure Lengend Snippet: A bioactivity assay of purified hFGF21 in a transfected NIH-3T3 cell line. The effect of the purified hFGF21 was determined in NIH-3T3 cells transfected with β-klotho. Dose-response proliferation curves of the cells were constructed from various concentrations of commercial hFGF21 from E. coli source and purified hFGF21.

Article Snippet: Commercial hFGF21 purified from inclusion bodies in E. coli (Cat# CYT-474) was acquired from Prospec (East Brunswick, NJ), and Dulbecco’s Modified Eagle’s Medium, foetal bovine serum, and penicillin-streptomycin from Gibco (MA).

Techniques: Purification, Transfection, Construct