recombinant human fgf21 rhfgf21  (PeproTech)

Journal: Frontiers in Pharmacology

Article Title: Sodium nitrate regulates senescence accompanied by aortic atherosclerosis in ApoE −/− mice through the miR-34a/FGF-21 axis

doi: 10.3389/fphar.2025.1562321

Figure Lengend Snippet: The sequences of the real-time PCR primers.

Article Snippet: FGF21 recombinant protein (50 ng/mL) (rhFGF21, HY-P7345, MedChemExpress, United States) were also used to treat HAoECs concomitantly with miR-34a mimic.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Frontiers in Pharmacology

Article Title: Sodium nitrate regulates senescence accompanied by aortic atherosclerosis in ApoE −/− mice through the miR-34a/FGF-21 axis

doi: 10.3389/fphar.2025.1562321

Figure Lengend Snippet: Effects of miR-34a and FGF21 in ApoE −/− mice and HAoECs. (A) Venn plots for GeneCards database search for senescence and AS-related miRNA. (B) Top 10 miRNAs with the highest correlation scores associated with senescence in the common 276 miRNAs. (C) QPCR was used to detect the expression level of miR-34a in ApoE −/− mice at 12 weeks and HAoECs at 24 h. n = 3. (D) QPCR analysis of FGF21 mRNA expression in ApoE −/− mice at 12 weeks and HAoECs at 24 h. n = 3. (E) QPCR was used to detect the expression levels of miR-34a in HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. n = 3. (F) QPCR analysis of FGF21 mRNA expression in HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. n = 3. (G) The CCK-8 method was used to evaluate the effect of the miR-34a mimic on the proliferation of HAoECs at 96 h. n = 3. (H) CCK-8 was used to detect the effect of the miR-34a inhibitor on the proliferation of HAoECs at 96 h. n = 3. (I) A representative image of SA-β-Gal-stained HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. n = 5. Bar = 100 μm. (J) Statistical analysis of the stained area in (I) . n = 5. (K) A representative image of γ-H2AX staining of HAoECs after treatment with the miR-34a mimic or inhibitor at 24 h. Bar = 20 μm. (L) Statistical analysis of the number of DNA damage foci in (K) . n = 5. (M) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in HAoECs after miR-34a mimic treatment at 24 h. n = 3. (N) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in HAoECs after miR-34a inhibitor treatment at 24 h. n = 3. (O) QPCR was used to detect the relative expression levels of the cell cycle-related genes p53, p21, p16 and Rb in HAoECs after miR-34a mimic treatment at 24 h. n = 3. (P) QPCR was used to detect the relative expression levels of the cell cycle-related genes p53, p21, p16 and Rb in HAoECs in different treatment groups after miR-34a-5p inhibitor treatment at 24 h. n = 3. (Q) Western blotting was used to detect the protein expression of p53, p21, and Rb in HAoECs after miR-34a mimic or inhibitor treatment at 24 h. n = 3. (R) Quantitative analysis of the results in (Q) . *p < 0.05, **p < 0.01, ***p < 0.001, vs. the NFD group or the control group or mimic-NC; ## p < 0.01, ### p < 0.001, vs. HFD group or H 2 O 2 group or inhibitor-NC.

Article Snippet: FGF21 recombinant protein (50 ng/mL) (rhFGF21, HY-P7345, MedChemExpress, United States) were also used to treat HAoECs concomitantly with miR-34a mimic.

Techniques: Expressing, CCK-8 Assay, Staining, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Sodium nitrate regulates senescence accompanied by aortic atherosclerosis in ApoE −/− mice through the miR-34a/FGF-21 axis

doi: 10.3389/fphar.2025.1562321

Figure Lengend Snippet: Effect of FGF21 on the senescence of HAoECs. (A) A CCK-8 kit was used to detect the effect of rhFGF21 on the proliferation of HAoECs at 48 h. (B) Statistical analysis of the HAoEC proliferation ability at 48 h (A) . n = 3. (C) Representative images of SA-β-Gal-stained HAoECs after treatment with the miRNA-34a mimic or rhFGF21 at 24 h. Bar = 100 μm. (D) Statistical analysis of the stained area in (C) . n = 5. (E) Representative images of γ-H2AX staining of HAoECs after miRNA-34a mimic and rhFGF21 treatment at 24 h. Bar = 20 μm. (F) Statistical analysis of the number of DNA damage foci in (E) . n = 5. (G) QPCR analysis of the mRNA expression of the inflammatory factors IL-1β, IL-6, IL-8 and TNF-α in HAoECs from different treatment groups at 24 h. n = 3. (H) QPCR was used to detect the relative mRNA expression of p53, p21, p16 and Rb in HAoECs after treatment with the miRNA-34a mimic or rhFGF21 at 24 h n = 3. (I) Western blotting was used to detect the protein expression of P53, P21, and RB in HAoECs after treatment with the miRNA-34a mimic or rhFGF21 at 24 h n = 3. (J) Quantitative analysis of the results in (I) . *p < 0.05, **p < 0.01, ***p < 0.001, vs. the mimic-NC group; # p < 0.05, ### p < 0.001, vs. the mimic group.

Article Snippet: FGF21 recombinant protein (50 ng/mL) (rhFGF21, HY-P7345, MedChemExpress, United States) were also used to treat HAoECs concomitantly with miR-34a mimic.

Techniques: CCK-8 Assay, Staining, Expressing, Western Blot

Journal: Science Advances

Article Title: Compromised macrophages contribute to progression of MASH to hepatocellular carcinoma in FGF21KO mice

doi: 10.1126/sciadv.ado9311

Figure Lengend Snippet: ( A ) Representative images of MASH-HCC histology by H&E and Oil Red O staining and immunohistochemistry detection of GF21 protein distribution in the regions of tumor (white dash circle) tissue and adjacent benign tissue. ( B ) Protein and mRNA levels of FGF21 by Western blotting and qPCR in the tumor and adjacent tissues. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. ( C ) Serum FGF21 protein levels by ELISA in the postresection patients with HCC (no relapse and relapse). ( D ) A cohort of 156 overweight (BMI ≥ 25 kg/m 2 ) patients with HCC from TCGA database were subgrouped into G1–2 patients and G3–4 patients to analyze the survival rate, based on high/low expression of FGF21 . ( E ) Representative images of Hyperion Imaging in paraffin-embedding tissues of patients with HCC. ( F to H ) Cell subpopulations of CD68 + macrophages, CD68 + CD163 + macrophages, and CD68 + CD163 + CD14 + macrophages in tumor and adjacent tissues. ( I ) the ratio of CD68 + CD163 + CD14 + macrophages in CD68 + macrophages in tumor and adjacent tissues. * P < 0.05 and ** P < 0.01.

Article Snippet: Treatments in animals were as follows: recombinant human FGF21 (rhFGF21) (no. 100-42; PeproTech) was injected subcutaneously every day at 125 μg/kg body weight.

Techniques: Staining, Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Imaging

Journal: Science Advances

Article Title: Compromised macrophages contribute to progression of MASH to hepatocellular carcinoma in FGF21KO mice

doi: 10.1126/sciadv.ado9311

Figure Lengend Snippet: ( A ) Volcano plot of DEGs (log2 fold change >1.5) by RNA-seq analysis in the isolated macrophages between FGF21KO-MASH mice and WT-CD controls. ( B ) Heatmap of the signature genes for monocytes and KCs by RNA-seq analysis between FGF21KO-MASH mice and WT-CD controls. ( C ) Running Enrichment Score of transcripts of genes of KCs/macrophages associated with fatty acid (FA) metabolism. ( D ) Heatmap of enzymes for FA metabolism by qPCR. Protein levels of FASN and CD36 by Western blotting in: ( E ) the WT/FGF21KO mice with WSHFD/CD diets, ( F ) the FGF21KO-MASH mice with rhFGF21 treatment compared to vehicle controls and WT-CD controls, and ( G ) FL83B/FL83B-FGF21KD cells treated with palmitic acid (PA), while 1% bovine serum albumin (BSA) was used as treatment control. ( H ) Protein levels of HSL and phosphorylated HSL at S563 and S660 by Western blotting in the WT/FGF21KO mice with WSHFD/CD diets. ( I ) Heatmap of long-chain FAs (LCFAs) detected by Waters ACQUITY UPLC Systems coupled with Waters Xevo TQ-S micro triple quadrupole mass spectrometer in the WT/FGF21KO mice with WSHFD/CD diets. 21KO, FGF21KO. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Treatments in animals were as follows: recombinant human FGF21 (rhFGF21) (no. 100-42; PeproTech) was injected subcutaneously every day at 125 μg/kg body weight.

Techniques: RNA Sequencing, Isolation, Western Blot, Control, Targeted Proteomics, Mass Spectrometry

Journal: Science Advances

Article Title: Compromised macrophages contribute to progression of MASH to hepatocellular carcinoma in FGF21KO mice

doi: 10.1126/sciadv.ado9311

Figure Lengend Snippet: ( A ) Protein levels of Alox15 by Western blotting in the liver tissues of WT/FGF21KO mice with WSHFD/CD diets. ( B ) Alox15 mRNA levels by qPCR analysis in the isolated hepatocytes and KCs and the protein levels of Alox15 by Western blotting in KCs from the WT/FGF21KO mice with WSHFD/CD diets. ( C ) Immunofluorescent staining using the antibodies of anti-ALOX15 and anti-F4/80 to detect the positive ALOX15 macrophages. Green, ALOX15-positive staining cells; red, F4/80-positive staining cells; blue, 4′,6-diamidino-2-phenylindole (DAPI) staining to detect the nuclei as a counterstain. ( D ) Protein levels of FAS and cleaved caspase-3 by Western blotting in the isolated mouse KCs challenged with 13-HODE and treated with N -acetyl- l -cysteine (NAC). ( E ) Protein levels of FAS and cleaved caspase-3 by Western blotting in the liver tissues from FGF21KO-MASH mice with rhFGF21 treatment compared to vehicle controls or WT-CD controls. ( F ) Histology-based NAFLD active score (NAS) in the WT/ALOX15KO mice with HFMCD/WSHFD diets. ( G ) CyTOF analysis in the isolated macrophages from mice to detect the KCs defined as F4/80 hi Tim4 + CX3CR1 − cells and the MoMFs defined as CD11b + Tim4 − CX3CR1 + cells in KC pool. The FGF21KO-MASH mice were treated daily with PD146176 or rhFGF21 for 4 weeks. ( H ) Analysis of the RNA-seq data for the mRNA fold changes of immune response signaling pathway and cytokine-cytokine interaction signaling. 21KO, FGF21KO. * P < 0.05 and ** P < 0.01.

Article Snippet: Treatments in animals were as follows: recombinant human FGF21 (rhFGF21) (no. 100-42; PeproTech) was injected subcutaneously every day at 125 μg/kg body weight.

Techniques: Western Blot, Isolation, Staining, RNA Sequencing

Journal: Science Advances

Article Title: Compromised macrophages contribute to progression of MASH to hepatocellular carcinoma in FGF21KO mice

doi: 10.1126/sciadv.ado9311

Figure Lengend Snippet: ( A ) Schematic diagram for establishing MASH-HCC models and the gross anatomy of tumor mass in four MASH-HCC models with CD, HFMCD, WSHFD, and HFD, respectively. ( B ) Maximal diameter of tumor nodules and number of tumor nodules in four MASH-HCC models. ( C ) Histology-based NAS in the MASH-HCC mice with HFMCD and WSHFD diets. ( D ) Schematic diagram for rhFGF21 treatment in the MASH-HCC mice with HFMCD and WSHFD diets. ( E ) CyTOF analysis of the KCs (F4/80 hi Tim4 + CX3CR1 − cells) and MoMFs (CD11b + Tim4 − CX3CR1 + cells) from the MASH mice and MASH-HCC mice, as well as the control mice. ( F ) LCFAs in the FGF21KO-MASH-HCC mice compared to normal WT controls. ( G ) Protein levels of cPLA2, Alox15, and MAGL by Western blotting in the liver tissues of FGF21KO-MASH-HCC mice compared to normal WT controls. ( H ) The serum FFA levels in FGF21KO-HCC mice in comparison with the WT-HCC mice, with three diets (CD, HFMCD, and WSHFD). ( I ) S1P levels in serum and liver tissue from FGF21KO-HCC mice in comparison with the WT-HCC mice, with three diets (CD, HFMCD, and WSHFD). 21KO, FGF21KO. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Treatments in animals were as follows: recombinant human FGF21 (rhFGF21) (no. 100-42; PeproTech) was injected subcutaneously every day at 125 μg/kg body weight.

Techniques: Control, Western Blot, Comparison

Journal: Science Advances

Article Title: Compromised macrophages contribute to progression of MASH to hepatocellular carcinoma in FGF21KO mice

doi: 10.1126/sciadv.ado9311

Figure Lengend Snippet: ( A ) Schematic diagram of S1P biosynthetic cascade enzymes with the fold changes (FGF21KO-preHCC versus WT-preHCC mice). ND, no detection; NC, no change; red, >2-fold up-regulated; blue, <2-fold down-regulated. ( B ) The S1P levels and protein levels of phosphorylated SPHK1 in liver issues from WT/FGF21KO-preHCC mice compared to that from WT/FGF21KO mice. ( C ) The protein levels of phosphorylated YAP and YAP in WT/FGF21KO-preHCC mice. ( D ) S1P content in FL83B/FL83B-FGF21KD cells and Hepal-6/Hepal-6-FGF21KD cells challenged with PA and treated with rhFGF21. ( E ) The protein levels of phosphorylated SPHK1 in FL83B/FL83B-FGF21KD cells and Hepal-6/Hepal-6-FGF21KD cells challenged with PA and treated with rhFGF21. ( F ) The protein levels of phosphorylated YAP in Hepal-6/Hepal-6-FGF21KD cells challenged with PA or PA + S1P and treated with SKI-II, an inhibitor of SPHK1. ( G ) The protein levels of phosphorylated YAP in Hepal-6/Hepal-6-FGF21KD cells challenged with S1P and treated with inhibitors of S1P receptors (JTE013 for S1PR2 and W146 for S1PR1). ( H ) Colony-forming assay to detect the number of cell colonies in Hepal-6 cells challenged with S1P and treated with JTE013 or verteporfin, an inhibitor of YAP. ( I ) Schematic diagram for JTE013 treatment in the MASH-HCC mice with WSHFD diets. ( J ) The protein levels of phosphorylated YAP in MASH-HCC mice with JTE013 treatment compared to vehicle controls. 21KO, FGF21KO. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Treatments in animals were as follows: recombinant human FGF21 (rhFGF21) (no. 100-42; PeproTech) was injected subcutaneously every day at 125 μg/kg body weight.

Techniques:

Journal: Cell reports

Article Title: Housing mice near vs. below thermoneutrality affects drug-induced weight loss but does not improve prediction of efficacy in humans

doi: 10.1016/j.celrep.2024.114501

Figure Lengend Snippet: Weight loss efficacy in DIO mice at 22°C and 30°C and in humans living with obesity

Article Snippet: Recombinant human FGF21 , Novo Nordisk A/S , Cat#NNC-0194-0000-0001.

Techniques:

Journal: Cell reports

Article Title: Housing mice near vs. below thermoneutrality affects drug-induced weight loss but does not improve prediction of efficacy in humans

doi: 10.1016/j.celrep.2024.114501

Figure Lengend Snippet: (A) Body weight during treatment of male DIO mice with human FGF21 (0.5 nmol/kg s.c. twice daily). (B) Body weight at day of transfer to the indirect calorimetry system (day −12) and at the end of treatment (day 18). (C and D) Fat mass and fat-free mass at days −12 and 18. (E) TEE during treatment. (F) Change in TEE vs. change in body weight. Changes were calculated by subtracting the mean of days 11 and 12 from the mean of days −1 and 0. (G) TEE (mean of days 11 and 12) vs. body weight (mean of days 11 and 12). Regression line and 95% confidence limits of the full experiment vehicle 22°C (light blue, n = 136, R 2 = 0.42, TEE = 0.3914•BW-5.824) and 30°C (pink, n = 136, R 2 = 0.29, TEE = 0.2407•BW-1.967) data. Days 11 and 12 were chosen due to the loss of TEE data from days 13–16. (H) RER (VCO 2 /VO 2 ). (I) Food intake. (J) Cumulative food intake (kcal) during treatment phase. (K) Physical activity. (L) Final physical activity level (average of days 15 and 16) vs. change in body weight. (M) Final physical activity EE data in (K) multiplied by body weight. Data are mean ± SEM. Biological replicates: n = 8/group, except n = 4/group for TEE and RER on days 13–16. Technical replicates: n = 1. p values compare the difference between days −12 and 18 by two-way ANOVA with uncorrected Fisher’s LSD post hoc testing within Ta or drug treatment. Color scheme: light blue, 22°C vehicle; pink, 30°C vehicle; dark blue, 22°C hFGF21; and red, 30°C hFGF21.

Article Snippet: Recombinant human FGF21 , Novo Nordisk A/S , Cat#NNC-0194-0000-0001.

Techniques: Activity Assay

Journal: Cell reports

Article Title: Housing mice near vs. below thermoneutrality affects drug-induced weight loss but does not improve prediction of efficacy in humans

doi: 10.1016/j.celrep.2024.114501

Figure Lengend Snippet: Primer information

Article Snippet: Recombinant human FGF21 , Novo Nordisk A/S , Cat#NNC-0194-0000-0001.

Techniques:

Journal: Cell reports

Article Title: Housing mice near vs. below thermoneutrality affects drug-induced weight loss but does not improve prediction of efficacy in humans

doi: 10.1016/j.celrep.2024.114501

Figure Lengend Snippet:

Article Snippet: Recombinant human FGF21 , Novo Nordisk A/S , Cat#NNC-0194-0000-0001.

Techniques: Recombinant, Activity Assay, Expressing, Software

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

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: Recombinant human FGF21 (CSB-AP002471HU, Cusabio Technology, LLC) was expressed in E. coli and harvested by ion exchange and hydrophobic interaction chromatography, as previously described ( ).

Techniques: Staining, Membrane