Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: GLN’s induction of FGF21 expression in the hippocampus, cortex, and striatum. After two weeks of GLN injection, the protein levels of FGF21 were measured in the hippocampus, cortex, and striatum of ND ( A ) or HFD mice ( B ) using Western blotting with β-actin as an internal control. The results represent the means ± S.E.M. ( n = 8). * p < 0.05 and *** p < 0.001 compared with the GLN 0 mg/mouse group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Injection, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: GLN-induced expression of FGF21 in hippocampal and striatal cells. HT22 hippocampal ( A ) and STHdh Q7/Q7 striatal cells ( B ) were treated with GLN (0, 1.25, 2.5, 5, 10, 20 mM) for 6 h for mRNA determination or 24 h for protein measurement. FGF21 mRNA was quantified using real-time PCR with GAPDH as an internal control, while FGF21 protein was examined through Western blotting with β-actin as an internal control. FGF21 production in the cultured medium was assessed with ELISA. The results represent the means ± S.E.M. ( n = 4). * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the GLN 0 mM group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: Attenuation of GLN-enhanced learning and memory functions and FGF21 production through FGFR1 inhibition in the hippocampus, hippocampal cells, and striatal cells. The ND-fed mice were administrated GLN (0, 30 mg/mouse) alone or in combination with PD173074 (2 mg/kg) through IP injection for 2 weeks, followed by the NORT ( A ). The FGF21 protein levels in the hippocampus ( B ) and in HT22 or STHdh Q7/Q7 cells treated with GLN (20 mM) alone or in combination with PD173074 (0.025, 0.1, 1 μM) for 24 h ( C ) were determined through Western blotting or ELISA. The results represent the means ± S.E.M. from 6 animals ( A ) or 4 animals ( B ) in each treatment group or from 4 independent experiments ( C ). * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the GLN 0 mg/mouse group or the GLN 0 mM group. # p < 0.05 and ### p < 0.001 compared with the GLN 30 mg/mouse or the GLN 20 mM only group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Inhibition, Injection, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: Illustration of the potential involvement of signaling molecules in GLN-mediated FGF21 production in hippocampal and striatal cells. ( A ) HT22 cells or ( B ) STHdh Q7/Q7 cells were pretreated with specific inhibitors targeting NF-κB (Parthenolide, 5 µM), MAPK p38 (SB203580, 20 µM), JNK (SP600125, 20 µM), ERK (PD98059, 20 µM), Akt (LY294002, 10 μM), PKA (H89, 5 µM), PKC (Bisindolylmaleimide, BIMI, 5 µM), PPARα (GW6471, 10 µM), or PPARγ (GW9662, 10 µM) for 1 h before the addition of GLN (20 mM) for an additional 24 h. The FGF21 concentration in the cultured medium was quantified using ELISA. The results represent the means ± S.E.M. ( n = 4). * p < 0.05 and *** p < 0.001 compared with the 0 mM group. # p < 0.05 and ### p < 0.001 compared with the GLN 20 mM group.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Glucosamine Enhancement of Learning and Memory Functions by Promoting Fibroblast Growth Factor 21 Production

doi: 10.3390/ijms25084211

Figure Lengend Snippet: Demonstration of the further induction of FGF21 through overexpression of NF-κB p65 in HT22 cells. HT22 cells were transfected with EGFP-p65 or EGFP plasmid and treated with GLN (20 mM) for 24 h. Western blotting was performed to detect the proteins for FGF21, GFP, EGFP-RelA, and p65 using β-actin as an internal control. The results represent the means ± S.E.M. ( n = 3). * p < 0.05 and *** p < 0.001 compared with the 0 mM group. && p < 0.01 between the EGFP and EGFP-RelA groups.

Article Snippet: The concentration of FGF21 in the culture medium was determined using an ELISA kit for mouse FGF21 (R&D systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.

Techniques: Over Expression, Transfection, Plasmid Preparation, Western Blot, Control

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: Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/jci94337

Figure Lengend Snippet: Figure 3. FGF21 secreted from pancreas induces OPC proliferation. (A) Quantitations of Fgf21 mRNA (left) and FGF21 protein (right) in individual organs of intact mice (n = 9); **P < 0.01. (B) Representative images of FGF21-immunolabeled pancreas of intact mice (n = 3). (C) Double IHC staining for FGF21 with the indicated cell markers in the pancreas of adult mice (n = 3). (D) BrdU incorporation in mouse OPCs after stimulation with serum from mice with FGF21 knockdown in the pancreas (n = 4); *P < 0.05, **P < 0.01, as determined by ANOVA with Tukey’s post hoc test. Error bars represent SEM. Scale bars: 50 μm (B); 10 μm (C).

Article Snippet: To assess the role of FGFs, we used recombinant mouse FGF15 (Abcam), recombinant mouse FGF21 (R&D Systems), and recombinant mouse FGF23 (R&D Systems).

Techniques: Immunolabeling, Immunohistochemistry, BrdU Incorporation Assay, Knockdown

Journal: Journal of Clinical Investigation

Article Title: Peripherally derived FGF21 promotes remyelination in the central nervous system

doi: 10.1172/jci94337

Figure Lengend Snippet: Figure 6. FGF21 promotes human OPC proliferation. (A) Representative image of β-klotho expression in an autopsied sample from healthy patient and a patient with multiple sclerosis. Graphs show quantitations as indicated in the images (n = 4 for healthy patients, 3 for multiple sclerosis patients); **P < 0.01. (B) BrdU incorporation in human OPCs after stimulation with recombinant FGF21 (n = 6 for control, 4 for FGF21); *P < 0.05 as determined by Student’s t test. Error bars represent SEM. Scale bar: 20 μm.

Article Snippet: To assess the role of FGFs, we used recombinant mouse FGF15 (Abcam), recombinant mouse FGF21 (R&D Systems), and recombinant mouse FGF23 (R&D Systems).

Techniques: Expressing, BrdU Incorporation Assay, Recombinant, Control

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: High UA levels stimulated HDAC6 and inhibited FGF21 expression in HUVECs. A, NO production detected by NO assay kit (n = 5). B, HDAC6 mRNA expression determined by RT-qPCR (n = 5). C, HDAC6 protein expression determined by WB analysis (n = 4). D, FGF21 mRNA level measured by RT-qPCR (n = 5). E, FGF21 protein level measured by WB analysis (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; # P < 0.05, ## P < 0.01, ### P < 0.001 versus 6 mg/dL UA.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: TSA attenuated UA-stimulated endothelial dysfunction in HUVECs. A, Representative cell morphology and ROS generation captured by a microscope (n = 3). B, TNF-α, IL-1β, and IL-6 mRNA levels determined by RT-qPCR (n = 5). C, NO production (n = 5). D, RT-qPCR analysis of CD31 and α-SMA (n = 5). E, WB analysis of CD31 and α-SMA (n = 4). F, α-SMA expression analyzed by immunofluorescence (n = 3). G, WB analysis of p-AKT/AKT and p-eNOS/eNOS (n = 4). H, RT-qPCR analysis of FGF21 (n = 5). I, WB analysis of FGF21 (n = 4). J, Level of histone H3 acetylation on the FGF21 promoter region, as analyzed by ChIP (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; # P < 0.05, ## P < 0.01, ### P < 0.001 versus 12 mg/dL UA; & P < 0.05, && P < 0.01 versus 12 mg/dL UA+TSA low.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Microscopy, Quantitative RT-PCR, Expressing, Immunofluorescence

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: FGF21 knockdown abrogated the effects of TSA on UA-induced inflammation, oxidative stress, endothelial dysfunction, and endothelial–interstitial transformation in HUVECs. A, RT-qPCR analysis of FGF21 (n = 3). B, RT-qPCR analysis of TNF-α, IL-1β, and IL-6 (n = 4). C, ROS generation (n = 3). D, NO production (n = 5). E, Levels of p-AKT/AKT and p-eNOS/eNOS measured by WB (n = 4). F, RT-qPCR analysis of CD31 and α-SMA (n = 4). G, WB analysis of CD31 and α-SMA (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; ### P < 0.001 versus 12 mg/dL UA; $$ P < 0.01, $$$ P < 0.001 versus 12 mg/dL UA+TSA high; & P < 0.05, && P < 0.01, &&& P < 0.001 versus 12 mg/dL UA+TSA high+NC.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Knockdown, Transformation Assay, Quantitative RT-PCR

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: LY294002 abrogated the effects of TSA on UA-induced vascular endothelial cell injury in HUVECs. A, NO production (n = 5). B, Levels of TNF-α, IL-1β, and IL-6 detected by RT-qPCR (n = 5). C, WB analysis of p-AKT/AKT, p-eNOS/eNOS, and p-FoxO3a/FoxO3a (n = 4). D, WB analysis of FGF21, CD31, and α-SMA (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; ## P < 0.01, ### P < 0.001 versus 12 mg/dL UA; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001 versus 12 mg/dL UA+TSA high; &&& P < 0.001 versus 12 mg/dL UA+TSA high+ LY294002.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Quantitative RT-PCR

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: Different from TSA, Mocetinostat, a class-Ⅰ selective HDAC inhibitor did not attenuate UA-stimulated endothelial dysfunction in HUVECs. A, HDAC1,2,3,6 activity detected by HDAC kits (n = 5). B, TNF-α, IL-1β, and IL-6 mRNA level detected by RT-qPCR (n = 5). C, NO production (n = 5). D, RT-qPCR analysis of FGF21 (n = 5). E, WB analysis of FGF21 (n = 4). F, Level of histone H3 acetylation on the FGF21 promoter region, as analyzed by ChIP (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; # P < 0.05, ## P < 0.01, ### P < 0.001 versus 12 mg/dL UA; $ P < 0.05, $$ P < 0.01, $$$ P < 0.001 versus 12 mg/dL UA+TSA high.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Activity Assay, Quantitative RT-PCR

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: HDAC6 knockdown alleviated UA-induced inflammation, oxidative stress, endothelial dysfunction, and endothelial–interstitial transformation in HUVECs. A, HDAC6 mRNA level measured by RT-qPCR (n = 3). B, Levels of TNF-α, IL-1β, and IL-6 detected by RT-qPCR (n = 5). C, HDAC6 activity (n = 5). D, NO production (n = 5). E, WB analysis of FGF21 (n = 4). F, WB analysis of p-AKT/AKT, p-eNOS/eNOS, and p-FoxO3a/FoxO3a (n = 4). G, WB analysis of CD31 and α-SMA (n = 4). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; ## P < 0.01, ### P < 0.001 versus 12 mg/dL UA; $$ P < 0.01, $$$ P < 0.001 versus 12 mg/dL UA+NC.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Knockdown, Transformation Assay, Quantitative RT-PCR, Activity Assay

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: TSA or TubA alleviated aortic inflammation and dysfunction in hyperuricemic mice. A, Blood pressure between the groups (n = 15). B, Serum UA (n = 7–8). C, Serum NO (n = 8). D, Serum FGF21 (n = 7–8). E, Serum TNF-α and IL-6 (n = 7–8). F, Levels of TNF-α and IL-6 mRNA in aorta tissue were determined by RT-qPCR (n = 5). G, Vasorelaxant responses to Ach and SNP in mice aortic rings precontracted with phenylephrine (n = 5). H, Representative images of mice aorta histopathology through H&E staining (magnification × 200) and used for analysis of media thickness. I, WB analysis of HDAC1, 2, 3, and 6 in aorta tissue (n = 5). J, WB analysis of p-AKT/AKT, p-eNOS/eNOS, and p-FoxO3a/FoxO3a in aorta tissue (n = 5). K, WB analysis of FGF21, CD31, and α-SMA in aorta tissue (n = 5). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001 versus CON; # P < 0.05, ## P < 0.01, ### P < 0.001 versus model; $ P < 0.05, $$ P < 0.01 versus TSA.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Quantitative RT-PCR, Histopathology, Staining

Journal: Journal of Cardiovascular Pharmacology

Article Title: HDAC Inhibitors Alleviate Uric Acid–Induced Vascular Endothelial Cell Injury by Way of the HDAC6/FGF21/PI3K/AKT Pathway

doi: 10.1097/FJC.0000000000001372

Figure Lengend Snippet: TSA or TubA alleviated renal inflammation and dysfunction in hyperuricemic mice. A, Serum BUN (n = 7–8). B, Serum CRE (n = 8). C, RT-qPCR analysis of TNF-α and IL-6 in renal tissues (n = 5). D, Representative images of mice renal tissue stained by H&E, Masson, and PAS (magnification × 200), and percentage of fibrotic area was analyzed. E, WB analysis HDAC1, 2, 3, and 6 (n = 5). F, WB analysis of p-AKT/AKT, p-eNOS/eNOS, and p-FoxO3a/FoxO3a (n = 5). G, WB analysis of FGF21, CD31, and α-SMA (n = 5). One-way ANOVA followed by Tukey's post hoc multiple-group comparisons. * P < 0.05, ** P < 0.001, *** P < 0.001 versus CON; # P < 0.05, ## P < 0.01, ### P < 0.001 versus model; $ P < 0.05, $$ P < 0.01 versus TSA.

Article Snippet: The serum levels of FGF21 (CSB-EL008627MO), TNF-α (CSB-E04741m), and IL-6 (CSB-E04639m) were determined using the enzyme-linked immunosorbent assay (ELISA) kits (CUSABIO life Sciences, Wuhan, China) according to the kit's protocols.

Techniques: Quantitative RT-PCR, Staining

Journal: EMBO Reports

Article Title: MAPL loss dysregulates bile and liver metabolism in mice

doi: 10.15252/embr.202357972

Figure Lengend Snippet: Representative immunoblots from liver extracts from three animals of each genotype: PERK auto‐phosphorylation along with the total protein and BIP and CHOP expressions are shown (*: unspecific signal). Representative immunoblots of eIF2α phosphorylation in liver extracts ( n = 3). mRNA expression by RNAseq of integrated stress response (ISR) genes in the liver of MAPL −/− mice. Fgf21 mRNA expression measured by qRT‐PCR performed on 3 different mouse tissues isolated from MAPL f/f and MAPL −/− males. N = 3 different mice per genotype. * P < 0.05, ** P < 0.01, *** P < 0.001 in an unpaired two‐tailed t ‐test. Representative immunoblots of increased FGF21 protein levels in liver from three pairs of mice from each strain. Serum FGF21 was quantified by ELISA ( n = 5 for each genotype, 2‐month‐old males). ** P < 0.01 in an unpaired two‐tailed t ‐test. Representative immunoblots from different tissues whole‐cell extracts from one male of each genotype: PERK auto‐phosphorylation, as well as BIP and CHOP expression are shown (Sto: stomach, *: remaining beta‐actin signal). Serum FGF21 was quantified by ELISA from tail vein injected adenoviral rescued mice ( n = 4 with two females and two males, n = 4 with two females and two males, n = 3 with two females and one male and n = 3 with one female and two males, for MAPL f/f + rtTA, MAPL −/− + rtTA, MAPL −/− + MAPL‐Flag and MAPL −/− + MAPL‐ΔRING‐Flag, respectively, 2–3 months old). * P < 0.05 in a one‐way ANOVA test. Liver Fgf21 gene expression by qRT‐PCR on livers of rescued mice (in triplicate, n = 7 with four females and three males, n = 7 with four females and three males, n = 5 with three females and two males and n = 6 with three females and three males, for MAPL f/f + rtTA, MAPL −/− + rtTA, MAPL −/− + MAPL‐Flag and MAPL −/− + MAPL‐ΔRING‐Flag, respectively, 2–3 months old). ** P < 0.01 in a Kruskal‐Wallis test. Representative immunoblot from rescued liver extracts probed for Fgf21, CHOP, P‐eIF2α and eIF2α as indicated (*: unspecific signal). Data are represented as mean ± SEM. Squares represent males whereas circles represent females. Source data are available online for this figure.

Article Snippet: Mitochondrial and peroxisomal anchored protein ligase was detected by rabbit polyclonal antibodies (HPA017681, 1:1,000, Sigma), PERK by rabbit polyclonal antibodies (100‐401‐962, 1:1,000, Rockland antibodies & assays), phospho‐PERK by rabbit monoclonal antibodies (3179, 1:1,000, Cell Signaling), eIF2α by mouse monoclonal antibodies (2103, 1:500, Cell Signaling), phospho‐eIF2α by rabbit polyclonal antibodies (SAB4504388, 1:500, Sigma), Fgf21 by goat polyclonal antibodies (AF3057, 1:500, R&D systems), CHOP by rabbit polyclonal antibodies (5554, 1:500, Cell Signaling), BiP by rabbit polyclonal antibodies (ADI‐SPA‐826‐D, 1:1,000, Enzo), CYP3A11 by rabbit polyclonal antibodies (13384, 1:500, Cell Signaling), CYP4A14 by goat polyclonal antibodies (sc‐46087, 1:500, Santa Cruz), CYP7A1 by rabbit polyclonal antibodies (ab65596, 1:500, Abcam), CYP27A1 by rabbit polyclonal antibodies (NBP2‐16061, 1:500, Novus Biologicals), SUMO1 by mouse monoclonal antibodies (332400, 1:1,000, Invitrogen), Hsp60 by mouse monoclonal antibodies (sc‐136291, 1:1,000, Santa Cruz), Hsp70 by rabbit polyclonal antibodies (ab137680, 1:1,000, Abcam), ABCD3 by mouse monoclonal antibodies (sab4200181, 1:1,000, Sigma), DRP1 by mouse monoclonal antibodies (611113, 1:1,000, BD Transduction Labs), Mfn2 by rabbit polyclonal antibodies (M6319, 1:1,000, Sigma), UCP1 by a polyclonal antibody (U6382, 1:500, Sigma), ACOX1 by a polyclonal antibody (10957‐1‐AP, 1:1,000, Proteintech), SCP2 by a polyclonal antibody (14377‐1‐AP, 1:1,000, Proteintech), PEX14 by a polyclonal antibody (ABC142, 1:1,000, Millipore), vinculin by a monoclonal antibody (V4505, 1:1,000, Sigma), β‐actin by rabbit polyclonal antibodies (SAB4502543, 1:1,000, Sigma) AIF by mouse monoclonal antibody (sc13116 1:1,000, SantaCruz), ABCB11 by polyclonal antibodies (PA527742 1:1,000, Thermo Fisher), EGFR by rabbit polyclonal antibodies (A00023 1:1,000, Boster), phospho‐EGFR by rabbit polyclonal antibodies (369700 1:1,000, Invitrogen) and β‐actin by mouse monoclonal antibodies (A2228, 1:1,000, Sigma).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Isolation, Two Tailed Test, Enzyme-linked Immunosorbent Assay, Injection

Journal: EMBO Reports

Article Title: MAPL loss dysregulates bile and liver metabolism in mice

doi: 10.15252/embr.202357972

Figure Lengend Snippet: Quantification of phospho‐PERK, CHOP and phospho‐eIF2α signals presented in Fig immunoblots ( n = 3 different mouse for each genotype). * P < 0.05, ** P < 0.01 in an unpaired two‐tailed t ‐test. Quantification of Fgf21 signals presented in Fig ( n = 3 for each genotype). Circulating and liver corticosterone levels were quantified as described in materials and methods ( n = 8 for each genotpye, 2‐month‐old males). *** P < 0.001 in an unpaired two‐tailed t ‐test. Quantification of FGF21, CHOP, phospho‐eIF2α and eIF2α signals presented in Fig ( n = 5, 5, 3 and 4 for MAPL f/f + rtTA, MAPL −/− + rtTA, MAPL −/− + MAPL‐Flag and MAPL −/− + MAPL‐ΔRING‐Flag, respectively, 2–3 months old). * P < 0.05, ** P < 0.01 in a Kruskal‐Wallis test. Representative immunoblot of CHOP protein expression of livers isolated from MAPL f/f and MAPL −/− animals injected or not with the empty virus rtTA. Data are represented as mean ± SEM. Squares represent males whereas circles represent females.

Article Snippet: Mitochondrial and peroxisomal anchored protein ligase was detected by rabbit polyclonal antibodies (HPA017681, 1:1,000, Sigma), PERK by rabbit polyclonal antibodies (100‐401‐962, 1:1,000, Rockland antibodies & assays), phospho‐PERK by rabbit monoclonal antibodies (3179, 1:1,000, Cell Signaling), eIF2α by mouse monoclonal antibodies (2103, 1:500, Cell Signaling), phospho‐eIF2α by rabbit polyclonal antibodies (SAB4504388, 1:500, Sigma), Fgf21 by goat polyclonal antibodies (AF3057, 1:500, R&D systems), CHOP by rabbit polyclonal antibodies (5554, 1:500, Cell Signaling), BiP by rabbit polyclonal antibodies (ADI‐SPA‐826‐D, 1:1,000, Enzo), CYP3A11 by rabbit polyclonal antibodies (13384, 1:500, Cell Signaling), CYP4A14 by goat polyclonal antibodies (sc‐46087, 1:500, Santa Cruz), CYP7A1 by rabbit polyclonal antibodies (ab65596, 1:500, Abcam), CYP27A1 by rabbit polyclonal antibodies (NBP2‐16061, 1:500, Novus Biologicals), SUMO1 by mouse monoclonal antibodies (332400, 1:1,000, Invitrogen), Hsp60 by mouse monoclonal antibodies (sc‐136291, 1:1,000, Santa Cruz), Hsp70 by rabbit polyclonal antibodies (ab137680, 1:1,000, Abcam), ABCD3 by mouse monoclonal antibodies (sab4200181, 1:1,000, Sigma), DRP1 by mouse monoclonal antibodies (611113, 1:1,000, BD Transduction Labs), Mfn2 by rabbit polyclonal antibodies (M6319, 1:1,000, Sigma), UCP1 by a polyclonal antibody (U6382, 1:500, Sigma), ACOX1 by a polyclonal antibody (10957‐1‐AP, 1:1,000, Proteintech), SCP2 by a polyclonal antibody (14377‐1‐AP, 1:1,000, Proteintech), PEX14 by a polyclonal antibody (ABC142, 1:1,000, Millipore), vinculin by a monoclonal antibody (V4505, 1:1,000, Sigma), β‐actin by rabbit polyclonal antibodies (SAB4502543, 1:1,000, Sigma) AIF by mouse monoclonal antibody (sc13116 1:1,000, SantaCruz), ABCB11 by polyclonal antibodies (PA527742 1:1,000, Thermo Fisher), EGFR by rabbit polyclonal antibodies (A00023 1:1,000, Boster), phospho‐EGFR by rabbit polyclonal antibodies (369700 1:1,000, Invitrogen) and β‐actin by mouse monoclonal antibodies (A2228, 1:1,000, Sigma).

Techniques: Western Blot, Two Tailed Test, Expressing, Isolation, Injection, Virus

Journal: EMBO Reports

Article Title: MAPL loss dysregulates bile and liver metabolism in mice

doi: 10.15252/embr.202357972

Figure Lengend Snippet:

Article Snippet: Mitochondrial and peroxisomal anchored protein ligase was detected by rabbit polyclonal antibodies (HPA017681, 1:1,000, Sigma), PERK by rabbit polyclonal antibodies (100‐401‐962, 1:1,000, Rockland antibodies & assays), phospho‐PERK by rabbit monoclonal antibodies (3179, 1:1,000, Cell Signaling), eIF2α by mouse monoclonal antibodies (2103, 1:500, Cell Signaling), phospho‐eIF2α by rabbit polyclonal antibodies (SAB4504388, 1:500, Sigma), Fgf21 by goat polyclonal antibodies (AF3057, 1:500, R&D systems), CHOP by rabbit polyclonal antibodies (5554, 1:500, Cell Signaling), BiP by rabbit polyclonal antibodies (ADI‐SPA‐826‐D, 1:1,000, Enzo), CYP3A11 by rabbit polyclonal antibodies (13384, 1:500, Cell Signaling), CYP4A14 by goat polyclonal antibodies (sc‐46087, 1:500, Santa Cruz), CYP7A1 by rabbit polyclonal antibodies (ab65596, 1:500, Abcam), CYP27A1 by rabbit polyclonal antibodies (NBP2‐16061, 1:500, Novus Biologicals), SUMO1 by mouse monoclonal antibodies (332400, 1:1,000, Invitrogen), Hsp60 by mouse monoclonal antibodies (sc‐136291, 1:1,000, Santa Cruz), Hsp70 by rabbit polyclonal antibodies (ab137680, 1:1,000, Abcam), ABCD3 by mouse monoclonal antibodies (sab4200181, 1:1,000, Sigma), DRP1 by mouse monoclonal antibodies (611113, 1:1,000, BD Transduction Labs), Mfn2 by rabbit polyclonal antibodies (M6319, 1:1,000, Sigma), UCP1 by a polyclonal antibody (U6382, 1:500, Sigma), ACOX1 by a polyclonal antibody (10957‐1‐AP, 1:1,000, Proteintech), SCP2 by a polyclonal antibody (14377‐1‐AP, 1:1,000, Proteintech), PEX14 by a polyclonal antibody (ABC142, 1:1,000, Millipore), vinculin by a monoclonal antibody (V4505, 1:1,000, Sigma), β‐actin by rabbit polyclonal antibodies (SAB4502543, 1:1,000, Sigma) AIF by mouse monoclonal antibody (sc13116 1:1,000, SantaCruz), ABCB11 by polyclonal antibodies (PA527742 1:1,000, Thermo Fisher), EGFR by rabbit polyclonal antibodies (A00023 1:1,000, Boster), phospho‐EGFR by rabbit polyclonal antibodies (369700 1:1,000, Invitrogen) and β‐actin by mouse monoclonal antibodies (A2228, 1:1,000, Sigma).

Techniques: