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: 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

Journal: Molecular Metabolism

Article Title: Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism

doi: 10.1016/j.molmet.2022.101534

Figure Lengend Snippet: Plasma FGF21 concentrations were associated with recent alcohol use phenotypes. Plasma FGF21 levels were positively correlated with recent alcohol use as determined by TLFB 90 days prior to blood collection i.e. (A) the number of drinking days, (B) the number of heavy drinking days, and (C) total drinks. (D) Plasma FGF21 levels were also positively correlated with blood levels of gamma-glutamyl transferase (GGT) which is commonly used as a marker for heavy alcohol use.

Article Snippet: We measured FGF21 levels in the iPSC-derived brain organoids and HepG2 cell lysates with an FGF21 ELISA kit (EK0994, Boster Biological Technology, Pleasanton, CA) in accordance with the manufacturer's instructions.

Techniques: Clinical Proteomics, Marker

Journal: Molecular Metabolism

Article Title: Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism

doi: 10.1016/j.molmet.2022.101534

Figure Lengend Snippet: (A) Schematic outline of our proteomics-informed genomics research strategy. (B–C) Q–Q plot and Manhattan plot for GWAS of plasma FGF21 levels. (D) The locus zoom plot displays that the top SNPs on chromosome 17 map within a gene cluster: CYGB , PRCD and SNHG16 . The SNP most highly associated with plasma FGF21 levels in patients with AUD was rs9914222 ( p: 4.6E-09).

Article Snippet: We measured FGF21 levels in the iPSC-derived brain organoids and HepG2 cell lysates with an FGF21 ELISA kit (EK0994, Boster Biological Technology, Pleasanton, CA) in accordance with the manufacturer's instructions.

Techniques: Clinical Proteomics

Journal: Molecular Metabolism

Article Title: Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism

doi: 10.1016/j.molmet.2022.101534

Figure Lengend Snippet: Biological functions of the rs9914222 SNP. (A) SNP-dependent plasma FGF21 levels in patients with AUD demonstrating that rs9914222 is an eQTL. (B) rs9914222 is associated with SNHG16 mRNA expression in several brain regions. https://gtexportal.org/home/snp/rs9914222 .

Article Snippet: We measured FGF21 levels in the iPSC-derived brain organoids and HepG2 cell lysates with an FGF21 ELISA kit (EK0994, Boster Biological Technology, Pleasanton, CA) in accordance with the manufacturer's instructions.

Techniques: Clinical Proteomics, Expressing

Journal: Molecular Metabolism

Article Title: Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism

doi: 10.1016/j.molmet.2022.101534

Figure Lengend Snippet: Biological functions of the SNHG16 gene. (A) FGF21 concentration was measured after the knockdown of SNHG16 using siRNA in HepG2 cells. Relative mRNA expression of SNHG16 and COMT was determined after the knockdown of SNHG16. COMT enzyme activity was also measured after the knockdown of SNHG16 in HepG2 cells. At least three independent experiments were performed. ∗ p < 0.05. (B) FGF21 concentration was determined before and after ethanol treatment of HepG2 cells. Relative mRNA expression of SNHG16 and COMT was determined in response to ethanol treatment. COMT enzyme activity was then measured using HepG2 cells treated with ethanol. ∗A p value ≤ 0.05 was considered statistically significant (two tailed paired t test). Three independent experiments were performed. All values are mean ± S.E.M.

Article Snippet: We measured FGF21 levels in the iPSC-derived brain organoids and HepG2 cell lysates with an FGF21 ELISA kit (EK0994, Boster Biological Technology, Pleasanton, CA) in accordance with the manufacturer's instructions.

Techniques: Concentration Assay, Knockdown, Expressing, Activity Assay, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism

doi: 10.1016/j.molmet.2022.101534

Figure Lengend Snippet: Ethanol induced FGF21 in iPSC-derived brain organoids and activated the release of catecholamines. (A) A schematic outline of procedures used during the differentiation of iPSC-derived brain organoids. The panel below the schematic displays representative examples of staining for tyrosin hydroxylase (TH), and Neuron-specific class III beta-tubulin (TUJ1). (B) the effect of ethanol on FGF21 concentration was measure using ELISA. Dopamine, norepinephrine, and epinephrine concentrations were measured using UPLC–Tandem Mass Spectrometry. (C) A schematic outline of the catecholamine biosynthesis pathway. (D) mRNA expression of SNHG16, MAOA, MAOB, COMT, TH, COMT, DDC, DBH, and PNMT in response to ethanol treatment (25 mM) for 7 days. Realtime PCR experiments were performed in iPSC-derived brain organoids (n = 3). The expression of these genes was determined after exposure to drug for 7 days. ∗A p value ≤ 0.05 was considered statistically significant (two tailed paired t test). All values shown are mean ± S.E.M.

Article Snippet: We measured FGF21 levels in the iPSC-derived brain organoids and HepG2 cell lysates with an FGF21 ELISA kit (EK0994, Boster Biological Technology, Pleasanton, CA) in accordance with the manufacturer's instructions.

Techniques: Derivative Assay, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Mass Spectrometry, Expressing, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism

doi: 10.1016/j.molmet.2022.101534

Figure Lengend Snippet: Gene expression of iPSC-derived brain organoids. (A) Schematic diagram illustrating the effects of ethanol on FGF21 which have implications for alcohol use. Specifically, FGF21 can be induced by ethanol. Our GWAS for plasma FGF21 showed that a SNP located 5′ of SNHG16 is associated with AUD. SNHG16 could regulate COMT expression and activity, which play a role in catecholamine metabolism. Finally, we determined that ethanol induced both FGF21 and catecholamines, including dopamine, norepinephrine and epinephrine using iPSC-derived brain organoids. (B – G) mRNA expression of SNHG16, TH, COMT, DDC, DBH, PNMT in response to drug treatment. Realtime PCR experiments were performed in iPSC-derived brain organoids (n = 3). The expression of those genes was determined after exposure to drugs for 7 days. ∗A p value ≤ 0.05 was considered statistically significant (two tailed paired t test). All values are mean ± S.E.M.

Article Snippet: We measured FGF21 levels in the iPSC-derived brain organoids and HepG2 cell lysates with an FGF21 ELISA kit (EK0994, Boster Biological Technology, Pleasanton, CA) in accordance with the manufacturer's instructions.

Techniques: Gene Expression, Derivative Assay, Clinical Proteomics, Expressing, Activity Assay, Two Tailed Test

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: Control

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, Control

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, Control

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, Control, 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: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Analysis of serum FGF19 and FGF21 levels in patients with viral or alcoholic hepatitis. Serum levels of FGF19 (A) and FGF21 (B) were determined by enzyme-linked immunosorbent assay. G1 (F0–F1, n=10), G2 (F2–F3, n=10), G3 (F4A–F4C, n=15). FGF, fibroblast growth factor. *p<0.01.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Analysis of FGF19, FGF21, and β-Klotho expression and inflammatory markers in liver tissue. Expression levels of mRNA for FGF19 (A), FGF21 (B), β-Klotho (C), IL-1β (D), IL-6 (E), and TNF-α (F) were determined by real-time polymerase chain reaction. G1 (F0–F1, n=10), G2 (F2–F3, n=10), G3 (F4A–F4C, n=15). FGF, fibroblast growth factor; IL, interleukin; TNF, tumor necrosis factor. *p<0.05, † p<0.01.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Correlation between the Fibrosis Stage and mRNA Expression of FGF19/21, β-Klotho, and Inflammatory Markers

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Expressing

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Effects of IL-1β on β-Klotho and FGF21 in Huh-7 cells. Huh-7 cells were incubated with IL-1β for 6 hours. β-Klotho expression levels (A and G), protein kinase B (B), extracelluar signal-regulated kinases (C), JNK (D), IκBα (E), and FGF21 (F and G) were determined by immunoblotting. IL, interleukin; FGF, fibroblast growth factor; JNK, c-Jun N-terminal kinase.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Incubation, Expressing, Western Blot

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: Signaling pathways that inhibit β-Klotho and induce FGF21 by IL-1β in Huh-7 cells. Huh-7 cells were pretreated with an NF-κB inhibitor (Bay11-7082, A and E), JNK inhibitor (SP600125, B and F), protein kinase B inhibitor (LY294002, C and G), or extracelluar signal-regulated kinases inhibitor (PD98059, D and H) for 20 minutes and then treated with 10 ng/mL of IL-1β for 6 hours to detect β-Klotho and FGF21. (I) Huh-7 cells were pretreated with 1 μM NF-κB inhibitor Bay11-7082, after which β-Klotho and FGF21 levels were determined by immunoblotting. (J) Huh-7 cells were pretreated with 0.5 μM JNK inhibitor SP600125, after which β-Klotho and FGF21 levels were determined by immunoblotting. (K) β-Klotho and FGF21 levels were assessed 6 hours after pretreatment with 1 μM NF-κB inhibitor Bay11-7082 in Huh-7 cells. (L) β-Klotho and FGF21 levels were assessed 6 hours after pretreatment with 0.5 μM JNK inhibitor SP600125 in Huh-7 cells. FGF, fibroblast growth factor; IL, interleukin; NF-κB, nuclear factor-κB; JNK, c-Jun N-terminal kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Protein-Protein interactions, Western Blot

Journal: Gut and Liver

Article Title: Expression of Fibroblast Growth Factor 21 and β-Klotho Regulates Hepatic Fibrosis through the Nuclear Factor-κB and c-Jun N-Terminal Kinase Pathways

doi: 10.5009/gnl17443

Figure Lengend Snippet: FGF21 inhibits IL-1β-induced growth retardation of hepatocytes. Huh-7 cells were incubated with 10 ng/mL of IL-1β (A), 500 ng/mL of FGF21 (B), or IL-1β+FGF21 (C), after which the expression of β-Klotho and PCNA was determined. (D) IL-1β-treated Huh-7 cells were incubated with or without FGF21 in a dose-dependent manner. Cell viability was measured by an methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. IL, interleukin; FGF, fibroblast growth factor; PCNA, proliferating cell nuclear antigen; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The lysates were subjected to Western blotting analysis under reducing conditions using previously validated human β-Klotho antibodies (R & D Systems), anti-phospho-Erk1/2, total Erk1/2, phospho-AKT, total AKT, phospho-JNK, and total JNK antibodies (Cell Signaling Technology, Danvers, MA, USA), phospho-IκBα, total IκBα, proliferating cell nuclear antigen (PCNA), and GAPDH antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and FGF21 antibody (Boster Biological Technology, Pleasanton, CA, USA).

Techniques: Incubation, Expressing, MTT Assay