Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Construction, purification, and characterization of FGF21-164. ( A ) FGF21-164 sequence and FGF21 sequence. In the FGF21-164 sequence, red marked the mutated amino acid and blue represents the TR acid sequence from hCD164. In the FGF21 sequence, the red-labeled amino acid represents the amino acid sequence whose n-terminal is deleted in FGF21-164. ( B ) Subcloned cell culture supernatants expressing FGF21-164 and FGF21 were analyzed using Western blotting (Lane M: molecular weight marker. Lane 1~7: FGF21-164 clones 1, 2, 3, A4, B2, F3, and H4. Line 8: FGF21 clone2). ( C ) The purified FGF21-164 and FGF21 were assayed using SDS-PAGE and Western blotting (Lane M: molecular weight marker. Lane 1: purified FGF21-164. Line 2: purified FGF21). ( D ) Purity and hydrodynamic radius of FGF21-164 were analyzed using SEC-HPLC. ( E ) MALDI-TOF mass-spectrometry analysis of FGF21-164. ([M + H] + denotes the singly charged ionic forms, and [M + 2H] 2+ denotes doubly charged ionic forms). ( F ) Circular Dichroism (CD) spectra of FGF21-164 and FGF21.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Purification, Sequencing, Labeling, Cell Culture, Expressing, Western Blot, Molecular Weight, Marker, Clone Assay, SDS Page, Mass Spectrometry, Circular Dichroism

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Glycan forms analysis of the protein FGF21-164. ( A ) O-linked glycan forms of FGF21-164. ( B ) N-linked glycan forms of FGF21-164.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of FGF21-164 in adipocytes induced by 3T3-L1. ( A ) The process of inducing 3T3-L1’s differentiation into adipocytes. ( B ) Reduction in glucose in medium stimulated by FGF21-164 in adipocytes induced by 3T3-L1. Data are means ± SEM (n = 6). * p < 0.05, *** p < 0.001, and **** p < 0.0001 compared to 0 μg/mL FGF21-164. ( C ) Phospho-Erk1/2-specific bands (Thr202/Tyr204; 44–42 kDa) were detected in 3T3-L1-derived adipocytes after FGF21-164 stimulation. ( D ) Evaluation of lipid droplet accumulation in adipocytes induced by 3T3-L1 with or without FGF21-164. The scale bar represents 50 μm.Data are the means ± SEM (n = 6). * p < 0.05, **** p < 0.0001.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Pharmacokinetics of FGF21-164. FGF21-164 plasma levels in C57BL/6 mice as determined via targeted LC-MS after i.v. (tail vein) bolus injection of FGF21-164 protein.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Liquid Chromatography with Mass Spectroscopy, Injection

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: The results of an oral glucose tolerance test applied to ob / ob mice after a single FGF21-164 treatment. ( A ) Blood glucose levels were measured at 0, 2, 3, 4, 5, 6, 7, and 8 h after the administration of FGF21-164. Data are the means ± SEM (n = 4–5). * p < 0.05 and ** p < 0.01 compared with the model. ( B ) The decrease in blood glucose levels post-administration relative to the baseline measurement at 0 h. Data are the means ± SEM (n = 4–5). * p < 0.05 compared with the model.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of a single administration of FGF21-164 on ob / ob mice. ( A , D , G ) Fasting glucose levels. Blood glucose levels were measured at 7, 21, and 28 days after a single dose of FGF21-164 was administered. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, ** p < 0.01, and **** p < 0.0001. ( B , E , H ) The OGTT at 7, 21, and 28 days. After oral glucose administration, blood glucose levels were measured. Data are the means ± SEM (n = 4–5). * p < 0.05, *** p < 0.001, and **** p < 0.0001 at 6 mg·kg −1 vs. model. # p < 0.05, ## p < 0.01 and #### p < 0.0001 at 12 mg·kg −1 vs. model. ( C , F , I ) The glucose AUC during the OGTT process. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, *** p < 0.001. ( J ) Body weight measured at 0, 7, 21, and 28 days. Data are the means ± SEM (n = 4–5). ns: no significance.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of repeated administration of FGF21-164. ( A ) Body weight changes. The DIO mice were treated with PBS or FGF21-164 once every 2 days. Data are the means ± SEM (n = 5–7). **** p < 0.0001 vs. Ctrl. #### p < 0.0001 for FGF21-164 vs. the model. ( B ) The liver sections were stained with Oil-red-O. The scale bar represents 100 μm. DIO mice showed many red lipid droplets in hepatic tissue (indicated by the black arrow), while lipid droplet levels were lower after the FGF21-164 treatment.

Article Snippet: Here, we describe a novel human FGF21 analog, designated as FGF21-164, which was generated through fusion protein technology and co-encoded by a mutated FGF21 gene and the TR gene of hCD164.

Techniques: Staining

Journal: International journal of molecular sciences

Article Title: Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP-Epac-CREB Signaling Pathway.

doi: 10.3390/ijms23105490

Figure Lengend Snippet: Figure 1. Pyruvate upregulated FGF21 expression and secretion in HepG2 cells. (A): pyruvate dose-dependently stimulated FGF21 gene expression after 12 h treatment in HepG2 cells (** p < 0.01 vs. control, n = 3). (B): FGF21 protein levels in cell medium significantly increased after pyruvate treatment (** p < 0.01 vs. control, n = 10). (C): The cell viability was not influenced by pyruvate treatment, as shown by MTT assay (n = 10). (D): D-LDH levels in cell medium were not changed by pyruvate treatment (n = 10).

Article Snippet: Human and mouse FGF21 ELISA kits, cAMP assay kits, the ALT activity assay kit, the AST activity assay kit and the LDH assay kit were obtained from Elabscience (Wuhan, China).

Techniques: Expressing, Gene Expression, Control, MTT Assay

Journal: International journal of molecular sciences

Article Title: Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP-Epac-CREB Signaling Pathway.

doi: 10.3390/ijms23105490

Figure Lengend Snippet: Figure 2. cAMP reduction caused pyruvate-stimulated FGF21 expression in HepG2 cells. (A): The ac- tivation of PPAR-α and AMPK was not involved in pyruvate-stimulated FGF21 expression (** p < 0.01 vs. control, n = 3). (B): AC activator forskolin, PDE inhibitor IBMX and 8-Bromo-cAMP administration significantly inhibited FGF21 expression and suppressed pyruvate-stimulated FGF21 expression (* p < 0.05 vs. control, # p < 0.05 vs. pyruvate group, n = 3). (C): Forskolin, IBMX and 8-Bromo-cAMP inhibited pyruvate-stimulated increase in FGF21 protein levels in cell medium (* p < 0.05 vs. control, # p < 0.05 vs. pyruvate group, n = 10). (D): Pyruvate decreased intracellular cAMP levels in HepG2 cells (** p < 0.01 vs. control, n = 12).

Article Snippet: Human and mouse FGF21 ELISA kits, cAMP assay kits, the ALT activity assay kit, the AST activity assay kit and the LDH assay kit were obtained from Elabscience (Wuhan, China).

Techniques: Expressing, Control

Journal: International journal of molecular sciences

Article Title: Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP-Epac-CREB Signaling Pathway.

doi: 10.3390/ijms23105490

Figure Lengend Snippet: Figure 4. Epac and CREB were involved in pyruvate-stimulated FGF21 expression in HepG2 cells. (A): Epac inhibitor ESI-09 but not PKA inhibitor H89 upregulated FGF21 expression and eliminated the stimulatory effect of pyruvate on FGF21 expression (** p < 0.01 vs. control, n = 3) (B): CREB inhibitor 666-15 upregulated FGF21 expression and eliminated the stimulatory effect of pyruvate on FGF21 expression (** p < 0.01 vs. control, n = 3). (C,D): Pyruvate reduced CREB phosphorylation without influencing the total CREB protein levels (** p < 0.01 vs. control, n = 3).

Article Snippet: Human and mouse FGF21 ELISA kits, cAMP assay kits, the ALT activity assay kit, the AST activity assay kit and the LDH assay kit were obtained from Elabscience (Wuhan, China).

Techniques: Expressing, Control, Phospho-proteomics

Journal: International journal of molecular sciences

Article Title: Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP-Epac-CREB Signaling Pathway.

doi: 10.3390/ijms23105490

Figure Lengend Snippet: Figure 5. Pyruvate upregulated FGF21 expression and secretion in mouse hepatic AML-12 cells. (A,B): Pyruvate stimulated FGF21 expression and secretion in AML-12 cells (* p < 0.05 and ** p < 0.01 vs. control, n = 6). (C): Pyruvate decreased intracellular cAMP levels in AML12 cells (* p < 0.05, n = 5). (D): Pyruvate increased PDE activities in AML-12 cells (* p < 0.05, n = 5).

Article Snippet: Human and mouse FGF21 ELISA kits, cAMP assay kits, the ALT activity assay kit, the AST activity assay kit and the LDH assay kit were obtained from Elabscience (Wuhan, China).

Techniques: Expressing, Control

Journal: International journal of molecular sciences

Article Title: Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP-Epac-CREB Signaling Pathway.

doi: 10.3390/ijms23105490

Figure Lengend Snippet: Figure 7. The diagram of pyruvate-stimulated FGF21 expression in hepatocytes. cAMP–Epac–CREB signaling inhibits FGF21 expression in human and mouse hepatocytes. Pyruvate activates PDEs to reduce cAMP levels and then inhibits cAMP–Epac–CREB signaling to upregulate FGF21 expression in hepatocytes.

Article Snippet: Human and mouse FGF21 ELISA kits, cAMP assay kits, the ALT activity assay kit, the AST activity assay kit and the LDH assay kit were obtained from Elabscience (Wuhan, China).

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: Serum FGF21 and its receptor KLB in humans and rats post-infarction. (A) Serum levels of FGF21 in patients with AMI, and ageand sex-matched healthy controls (n=50). (B) Serum levels of FGF21 in rats post-infarction (n=5). (C) Relative mRNA levels of KLB receptors in heart tissue (n=5). (D) Relative protein levels of KLB, FGF21 and α-klotho in heart tissue (n=5). (E) Representative images of echocardiography (n=5). (F) Measurements of LVEF (n=5). (G) Measurements of LVFS (%) (n=5). (H) Spearman's correlation analyses between CK-MB and serum levels of FGF21 (n=30). (I) Spearman's correlation analyses between cTnT and serum levels of FGF21 (n=30). (J) Spearman's correlation analyses between cTnT and KLB levels (n=30). ** P<0.01 and *** P<0.001 vs. the control (Con) group; ns, not significant. KLB, β-klotho; FGF21, fibroblast growth factor 21; AMI, acute myocardial infarction; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: KLB overexpression enhances the protective effects of FGF21 treatment on hypoxia-induced cardiomyocyte injury in vitro . (A) Viability of H9C2 cells exposed to hypoxia (HX), FGF21 and KLB overexpression (n=5). (B) Representative images of DHE staining (scale bar, 100 μ m), and DHE intensity was quantified to reflect ROS levels. (C) Representative TUNEL staining images of KLB-overexpressing H9C2 cells exposed to hypoxia and treated with FGF21 (n=4; scale bar, 100 μ m). (D) LDH leak from KLB-overexpressing H9C2 cells exposed to hypoxia and treated with FGF21 (n=8). (E) JC-1 staining of H9C2 cells (scale bar, 50 μ m). Mitochondrial membrane potential was estimated by the ratio of JC-1 aggregates (red, healthy mitochondria) and JC-1 monomers (green, depolarized mitochondria, n=5). ** P<0.01 and *** P<0.001 vs. the control (Con) group; ## P<0.01 and ### P<0.001 vs. hypoxia (HX). KLB, β-klotho; FGF21, fibroblast growth factor 21; AMI, acute myocardial infarction; LDH, lactate dehydrogenase.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques: Over Expression, In Vitro, Staining, TUNEL Assay, Membrane

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: Assessment of plasmid@CMB preparation and delivery efficiency via UTMD technology. (A) The preparation of CMB particles. (B) Size and potential distribution of the CMBs. (C) The binding rates of several doses of plasmid on CMBs (n=4). (D) Representative images of KLB@CMBs. KLB plasmid labeled by PI staining (red) and the outline of CMB (bright) are shown (scale bar, 50 μ m). (E) The binding rates of KLB@CMBs were assessed using flow cytometry (n=4). (F) Typical ultrasound contrast images of KLB@CMBs in the heart before injection, at ultrasound-targeted CMB blast, and after the CMB blast. After the CMB injection, microbubbles with high-echo intensity filled the ventricle chambers and wall tissue. The second harmonic mode with an electrocardiograph-mediated trigger was applied to activate microbubble bursting, and then only a small number of microbubbles remained and showed a low echo shadow. (G) Fluorescence images of GFP expression after plasmid containing GFP gene delivered by UTMD technique (DAPI for nucleus; green for GFP; scale bar, 50 μ m). and *** P<0.001 vs. the control (Con) group; and ### P<0.001 vs. 20 μ g plasmid. CMB, cationic microbubble; UTMD, ultrasound-targeted microbubble destruction; KLB, β-klotho; FGF21, fibroblast growth factor 21.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques: Plasmid Preparation, Binding Assay, Labeling, Staining, Flow Cytometry, Injection, Fluorescence, Expressing

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: KLB@CMBs amplify FGF21 treatment, improving heart dysfunction and the infarction area in rats post-AMI. (A) UTMD mediated the overexpression of cardiac KLB in rats post-infarction repeated at one-day intervals 3 times. *** P<0.001 vs. the control (Con) group. (B) Representative images of echocardiography at 4 weeks after AMI surgery. (C) The LVEF and LVFS (n=5-6). (D) Representative hematoxylin and eosin-stained images of heart sections (scale bar, 100 μ m). (E) Injury scores assessed according to hematoxylin and eosin images (n=5-6). (F) Representative images of Masson's staining were conducted to assess myocardial fibrosis. (G) The relative area of myocardial fibrosis was estimated according to Masson's staining (n=5-6). ** P<0.01 and *** P<0.001 vs. AMI; ## P<0.01 and ### P<0.001 vs. the AMI + FGF21 group. AMI, acute myocardial infarction; KLB, β-klotho; FGF21, fibroblast growth factor 21; CMB, cationic microbubble; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques: Over Expression, Staining

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: The combination of KLB@CMBs and FGF21 treatment inhibits myocardial oxidative stress compared with the use of FGF21 alone in rats post-infarction. (A) DHE staining in the heart sections (scale bar, 100 μ m). (B) Western blot analysis and quantification of KLB expression in the heart post-infarction with or without KLB delivery and FGF21 administration. (C) ATP generation in heart tissues. *** P<0.001 vs. AMI; ## P<0.01 and ### P<0.001 vs. the AMI + FGF21 group. AMI, acute myocardial infarction; KLB, β-klotho; FGF21, fibroblast growth factor 21; CMB, cationic microbubble.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques: Staining, Western Blot, Expressing

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: Cardiac delivery of KLB enhances the antioxidant effects of FGF21 in the heart following AMI. (A) Western analysis and quantification of Nrf2 and KEAP1 expression in H9C2 cells (n=4). (B) The protein expression of HO-1, NQO1, Gstp1 and GCLM was assessed using western blot analysis (n=4). (C) Representative images of echocardiography at 4 weeks following AMI surgery. (D and E) The LVEF and LVFS were calculated (n=5). (F and G) DHE staining of the heart section (scale bar, 50 μ m) and quantification (n=5). ** P<0.01 and *** P<0.001 vs. AMI; # P<0.05, ## P<0.01 and ### P<0.001 vs. the AMI + KLB@cMBs + FGF21 group; ns, not significant. AMI, acute myocardial infarction; KLB, β-klotho; FGF21, fibroblast growth factor 21; CMB, cationic microbubble; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening; Nrf2, nuclear factor erythroid 2-related factor 2; KEAP1, kelch-like ECH-associated protein 1; HO-1, heme oxygenase 1; NQO1, NAD(P)H quinone dehydrogenase 1; Gstp1, glutathione S-transferase pi-1; GCLM, glutamate-cysteine ligase modifier subunit.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques: Western Blot, Expressing, Staining

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: Mitochondrial quality in the heart is improved by the UTMD-mediated KLB delivery and FGF21 treatment in rats post-infarction. (A) JC-1 staining of heart section post-infarction (scale bar, 100 μ m). (B) The ratio of JC-1 aggregates (red) and monomers (green) was used to assess mitochondrial membrane potential (n=5-6). (C) Mitochondrial OCR profile in H9C2 cells using an XF24 Extracellular Flux Analyzer. Oligomycin (1 μ mol/l), FCCP (4 μ mol/l) and rotenone (0.5 μ mol/l) plus antimycin A (0.5 μ mol/l) were added sequentially. (D) FCCP-related respiration, (E) basal respiration, (F) maximal respiration and (G) ATP turnover were calculated (n=4). *** P<0.001 vs. AMI; ## P<0.01 and ### P<0.001 vs. the AMI + KLB@CMBs + FGF21 group. CMB, cationic microbubble; UTMD, ultrasound-targeted microbubble destruction; KLB, β-klotho; FGF21, fibroblast growth factor 21; OCR, oxygen consumption rate; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; AMI, acute myocardial infarction; HX, hypoxia.

Article Snippet: The serum levels of creatine kinase-myocardial band (CK-MB; E-EL-R1327c, Elabscience Biotechnology Co., Ltd.), cardiac troponin T (cTnT; E-EL-R0151c Elabscience Biotechnology Co., Ltd.), KLB (ml028418, Mlbio) and FGF21 (CSB-EL008627RA, Cusabio Biotechnology Co., Ltd.) were measured according to the instructions provided with the kits.

Techniques: Staining, Membrane

Journal: Food Science & Nutrition

Article Title: The protective effect of sulforaphane on type II diabetes induced by high‐fat diet and low‐dosage streptozotocin

doi: 10.1002/fsn3.2040

Figure Lengend Snippet: Effects of different dosages of SFN on serum FGF21 and LPS level in type II diabetes. (a) Serum FGF21 level. (b) Serum LPS level. Value is the mean ± SEM ( n = 8). Data not sharing a common superscript differ significantly among groups ( p < .05, ANOVA)

Article Snippet: Insulin, lipopolysaccharide (LPS), and fibroblast growth factor21 (FGF21) were measured according to the commercial kits (Elabscience Biotechnology, WuHan, Hubei, China).

Techniques:

Journal: Scientific Reports

Article Title: Administration of Danhong Injection to diabetic db/db mice inhibits the development of diabetic retinopathy and nephropathy

doi: 10.1038/srep11219

Figure Lengend Snippet: After treatment, liver, white adipose tissue (WAT), muscle and blood samples were collected to prepare total cellular protein, total RNA, total lipid and serum, respectively. Expression of PCK1, G6Pase and GCK ( A ); AMPKα and pi-AMPKα ( B ); IRS1, FGF21 and PPARγ ( C , D ) protein was determined by Western blot, respectively. *: vs. control db/db mice, p < 0.05 (n = 3); Expression of FGF21 [E, *: vs. wild type mice; **: vs . control db/db mice, p < 0.05 (n = 3)], PPARγ [F, *: vs. control db/db mice, p < 0.05 (n = 3)], ATGL, HSL, CPT1α, LCAD, MCAD, ACOX1, Cyto-C, ATPase and PPARα [G, *: vs. control db/db mice, p < 0.05 (n = 3)] mRNA was determined by real time RT-PCR, respectively; Serum FGF21 levels [E, *: vs. wild type mice; **: vs . control db/db mice, p < 0.05 (n = 5)] and liver TG content [F, *: vs. wild type mice, p < 0.05 (n = 5)] were determined using assay kits, respectively.

Article Snippet: Serum samples were also used to determine FGF21 levels using a mouse FGF21 ELISA kit (Boster, Wuhan, China) according to the manufacture’s instruction.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Administration of Danhong Injection to diabetic db/db mice inhibits the development of diabetic retinopathy and nephropathy

doi: 10.1038/srep11219

Figure Lengend Snippet: Sequences of the primers for real time RT-PCR analysis.

Article Snippet: Serum samples were also used to determine FGF21 levels using a mouse FGF21 ELISA kit (Boster, Wuhan, China) according to the manufacture’s instruction.

Techniques: Quantitative RT-PCR