Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: Circulating adiponectin levels and risk of nasopharyngeal carcinoma in retrospective and prospective cohorts

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques:

Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: Adiponectin suppresses nasopharyngeal carcinoma growth. A , B 1 × 10 6 CNE-2 cells were injected subcutaneously into 5- to 6- week-old adiponectin-deficient nude mice, or the control nude mice ( n = 6 per group). Tumor growth were monitored by measuring the tumor volume for 10 days. Next, the mice were sacrificed, and tumors were collected, measured, weighed. C The plate colony assay was performed to determine colony-formation ability. CNE-2 and C666-1 cells were treated with various concentrations of adiponectin for 7 days. Graphs show the number of colonies. D EdU incorporation assay was performed to determine cell proliferation. CNE-2 and C666-1 cells were treated with various concentrations of adiponectin for 48 h. Bars: 50 μm. Graphs show the relative cell proliferation percentage. E , F CNE-2 and C666-1 cells were incubated with adiponectin (40 μg/mL) for 48 h. At the end of incubation, the cells were collected for FACS analysis. G Western blot analysis of p-AMPKα (T172), p-LKB1 and p-ERk1/2 in cultured CNE-2 and C666-1 cells after the treatment with adiponectin (40 μg/mL) for 30 min. Results are presented as mean ± SD of three independent experiments performed in triplicate. * P < 0.05, ** P < 0.01, *** P < 0.001, # P < 0.0001

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Injection, Colony Assay, Incubation, Western Blot, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: Adiponectin suppresses proliferation of NPC cells via AMPK activation. A , B CNE-2 and C666-1 cells were pretreated with compound C (10 mM) followed by treatment with adiponectin (40 μg/mL) for 24 h. Cell cycle was then analyzed using flow cytometer. # P < 0.05 and ## P < 0.01 compared to cells treated with adiponectin but not ComC; ** P < 0.01 compared with cells treated without adiponectin and ComC. C CNE-2 and C666-1 cells viability was determined after treatment with or without adiponectin for 48 h in the presence or absence of ComC (10 μM). ** P < 0.01, *** P < 0.001. D CNE-2 and C666-1 cells were pretreated with compound C (10 mM), p-AMPKα (Thr172) protein level was then determined by Western blot analysis after the treatment with adiponectin (40 μg/mL) for 30 min; cyclin D1, p21, and p27 protein level was then determined by Western blot analysis after the cells were exposed to adiponectin (40 μg/mL) for 48 h. E Quantitative analysis of p-AMPKα level was performed by densitometric analysis. Results are presented as mean ± SD of three independent experiments performed in triplicate. ** P < 0.01, *** P < 0.001

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Activation Assay, Flow Cytometry, Western Blot

Journal: Journal of Translational Medicine

Article Title: Adiponectin suppresses tumor growth of nasopharyngeal carcinoma through activating AMPK signaling pathway

doi: 10.1186/s12967-022-03283-0

Figure Lengend Snippet: AdipoR1 and AdipoR2 mediate the anti-proliferative effect of adiponectin in NPC cells. A , B Expression of AdipoR1 and AdipoR2 was determined by qRT-PCR and Western blot in NPC cell lines. C , D CNE-2 and C666-1 cells were transfected with 50 μM siRNAs of NC, AdipoR1, or AdipoR2. The relative amounts of each AdipoR1/R2 mRNA against β-actin were measured with qRT-PCR. E Effect of the knockdown of AdipoR1 and AdipoR2 expression on the cell viability of CNE-2 and C666-1 cells treated with or without 40 μg/mL adiponectin for 48 h. F CNE-2 and C666-1 cells were transfected with AdipoR1, AdipoR2 siRNA or NC siRNA and treated with 40 μg/mL adiponectin for 30 min. AMPKα and p-AMPKα (T172) protein levels were determined by Western blot analysis. Quantitative analysis of p-AMPKα level was performed by densitometric analysis and shown in the below part. Results are presented as mean ± SD of three independent experiments performed in triplicate. ** P < 0.01, *** P < 0.001

Article Snippet: Recombination human full-length adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

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

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: Abbreviations

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques:

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: Structural overview of CTRP3. (A) Two splice variants were identified and designated as CTRP3A and CTRP3B. (B) Organization of the CTRP3A and CTRP3B genes and proteins. The human CTRP3A gene is 25.3 kb in size, consists of six exons and five introns, and is located on chromosome 5p13. Exons 1, 2, 3, 4, 5, and 6 of the CTRP3A gene are 171, 112, 155, 130, 100, and 2885 bp in size, respectively. The size of each intron is also indicated. Exon 1B (gray square) contains a 219-nucleotide sequence found in CTRP3B cDNA, coding for an extra 73 amino acid residues. A potential Nlinked glycosylation site is circled. The consensus splice donors are shown in italic type. This figure was originally published in The Journal of Biological Chemistry (51) and image reproduced according to the copyright policy of the ASBMB. © the American Society for Biochemistry and Molecular Biology.

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: Sequencing, Glycoproteomics

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: Complete Summary of the In Vitro Functions of CTRP3

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: In Vitro, Produced, Phospho-proteomics, Migration, Knockdown, Recombinant, Expressing, Control, Activity Assay, Cell Culture, In Vivo, Over Expression, Transfection

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: CTRP3 Expression In Vitro

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: Expressing, Control, Activity Assay, Blocking Assay, Activation Assay, Phospho-proteomics, Infection, Knockdown

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: Metabolic Effects of CTRP3 In Vivo

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: Knock-Out, Transgenic Assay, Over Expression, Injection, Recombinant, Suspension, Clinical Proteomics

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: Cross-Sectional Studies Regarding CTRP3 Levels

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: Control, Filtration, Biomarker Discovery

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: Summary of Cardiovascular Effects of CTRP3 Treatment

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: Produced, Migration, Recombinant, Knockdown, Phospho-proteomics, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Comprehensive Physiology

Article Title: C1q/TNF-Related Protein 3 (CTRP3) Function and Regulation

doi: 10.1002/cphy.c160044

Figure Lengend Snippet: CTRP3 and Inflammation

Article Snippet: Rat vascular smooth muscle cells (VSMCs) , Feng et al. 2016 ( 15 ) , Recombinant human globular CTRP3 (Aviscera Bioscience, no catalog # provided), or siRNA knockdown of CTRP3 , CTRP3 increase ATP synthesis and oxidative phosphorylation complex proteins, reversed with knockdown. CTRP3 increases mitochondrial ROS.

Techniques: Recombinant, Expressing, Control, Transgenic Assay, Over Expression, Knock-Out, Injection, IV Injection

Journal: Oncotarget

Article Title: Adiponectin receptor agonists inhibit leptin induced pSTAT3 and in vivo pancreatic tumor growth

doi: 10.18632/oncotarget.19905

Figure Lengend Snippet: Human and murine normal and pancreatic cancer samples were immunostained for ADIPOR1 and ADIPOR2. (A) strong staining is present in the acinar tissue of human normal samples for both receptors but is decreased in the human pancreatic tumor samples. (B) murine pancreatic cancer tissue samples from PKT mice demonstrate a high level of staining in the adjacent acinar tissue and a reduced staining in the dysplastic pancreatic tumor. Graphs represent quantitative analysis of mean fluorescence from single channel images. Scale bar is 100μm. (C-D) quantitative real time PCR analysis was used to determine the relative expression of AdipoR1 (white bars) and AdipoR2 (black bars) in human and murine pancreatic tumor samples (PDAC) as well as in pancreatic cancer cell lines, relative to the level expressed in the normal pancreas (NP) tissue. (E) quantitative real time PCR analysis for adiponectin expression in human and mouse pancreatic cancer cell lines and mouse tissue, relative to human adipose tissue (AT), 3T3L1 cells differentiated to adipocytes (3T3L1) and mouse AT, respectively. Statistical analysis was performed separately for ADIPOR1 and ADIPOR2 protein levels (unpaired t-test) as well as AdipoR1 and AdipoR2 expression (two-way Anova), or adiponectin expression (ordinary one-way Anova) (*P ≤ 0.05, ***P ≤ 0.001, ****P ≤ 0.0001).

Article Snippet: Recombinant globular adiponectin (1688-AC or 1119-AC), full length adiponectin (1065-AP or 5095-AC), and Leptin (Mouse 498-OB or human 398-LP) from R&D System was used for in vitro analysis.

Techniques: Staining, Fluorescence, Real-time Polymerase Chain Reaction, Expressing

Journal: Oncotarget

Article Title: Adiponectin receptor agonists inhibit leptin induced pSTAT3 and in vivo pancreatic tumor growth

doi: 10.18632/oncotarget.19905

Figure Lengend Snippet: (A) recombinant globular adiponectin (gADN) at 1μg/mL or full length adiponectin (fADN) at 10μg/mL were incubated with human (Panc1 and MiaPaca2) and mouse (Panc02) pancreatic cancer cell lines, labeled with EdU and measured for active proliferation. DMSO was used as a control. Statistical analysis was performed using unpaired t-test (***P ≤ 0.001, ****P ≤ 0.0001). (B) human (Panc1 and MiaPaca2) and mouse PDAC cells (Panc02, P-4313 and K-8484) were treated with increasing concentration of AdipoRon for 48h before measuring proliferation via EdU, which showed dose-dependent inhibition for each PDAC cell line. Statistical significance was determined using ordinary one-way Anova (*P ≤ 0.05, ****P ≤ 0.0001).

Article Snippet: Recombinant globular adiponectin (1688-AC or 1119-AC), full length adiponectin (1065-AP or 5095-AC), and Leptin (Mouse 498-OB or human 398-LP) from R&D System was used for in vitro analysis.

Techniques: Recombinant, Incubation, Labeling, Control, Concentration Assay, Inhibition

Journal: Oncotarget

Article Title: Adiponectin receptor agonists inhibit leptin induced pSTAT3 and in vivo pancreatic tumor growth

doi: 10.18632/oncotarget.19905

Figure Lengend Snippet: Murine P-4313 and K-8484 cancer cells were orthotopically injected into syngeneic wildtype mice and allowed to grow for two weeks, after which mice were treated with vehicle or AdipoRon 5mg/kg/day for 2 weeks. (A) at endpoint, AdipoRon treated mice had a significantly reduced tumor size. (B) tumor proliferation measured by Ki67 positive tumor area per tumor section was significantly reduced after AdipoRon treatment. (C) no significant difference was detected in relative expression of AdipoR1 (white bars) and AdipoR2 (black bars) by quantitative real time PCR analysis between control and AdipoRon treated tumors. (D) ELISA analysis showed no significant difference in circulating serum adiponectin levels between the two groups of mice. (P-4313 filled symbols and K-8484 open symbols). Statistical analysis was performed using unpaired t-test (*P ≤ 0.05, **P ≤ 0.01).

Article Snippet: Recombinant globular adiponectin (1688-AC or 1119-AC), full length adiponectin (1065-AP or 5095-AC), and Leptin (Mouse 498-OB or human 398-LP) from R&D System was used for in vitro analysis.

Techniques: Injection, Expressing, Real-time Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Drosophila Adiponectin Receptor in Insulin Producing Cells Regulates Glucose and Lipid Metabolism by Controlling Insulin Secretion

doi: 10.1371/journal.pone.0068641

Figure Lengend Snippet: (A) Amino acid sequence comparison between dAdipoR and human adiponectin receptor 1. Seven transmembrane domain regions are marked by upper lines. The putative interaction residues with the adiponectin are marked in blue. (B-I) Drosophila brains from the larva (B-E) and the adult (F-I) were immunostained with the dAdipoR antibody (green) and the IPCs marker, Dilp2>DsRed (red). dAdipoR staining was detected in IPCs (dot boxes), SOG neurons (arrows), and lateral neurons (arrowheads). Larval IPCs in B was enlarged in C-E and adult IPCs in F was enlarged in G-I, showing that dAdipoR staining in IPCs was overlapped with the IPCs marker. Scale bars are 100 µm (B, F) and 20 µm (E, I).

Article Snippet: The recombinant human globular adiponectin was purchased from R&D Systems.

Techniques: Sequencing, Comparison, Marker, Staining

Journal: PLoS ONE

Article Title: Drosophila Adiponectin Receptor in Insulin Producing Cells Regulates Glucose and Lipid Metabolism by Controlling Insulin Secretion

doi: 10.1371/journal.pone.0068641

Figure Lengend Snippet: (A) The dose effect of human adiponectin on Dilp2 secretion measured by Dilp2 staining intensity. 10 to 20 µg/ml of human adiponectin significantly induced Dilp2 secretion. (B-F) Images and relative intensities of Dilp2 staining in larval IPCs after treating with human adiponectin (10 µg/ml). DIlp2 secretion induced by human adiponectin was inhibited in IPCs of Dilp2>dAdipoR-Ri larvae. Data are presented as means ±SEM; **p<0.01. Scale bar is 20 µm (F).

Article Snippet: The recombinant human globular adiponectin was purchased from R&D Systems.

Techniques: Staining

Journal: bioRxiv

Article Title: The hepatokine FGL1 regulates hepcidin and iron metabolism during the recovery from hemorrhage-induced anemia in mice

doi: 10.1101/2023.04.06.535920

Figure Lengend Snippet: Relative HAMP expression in Hep3B and HepG2 cells in response to BMP6 (25 ng/ml, 6h) (A) or BMP6 + recombinant FGL1 (10µg/ml) (B). Relative ID1 expression in hepatoma cell lines treated with BMP6 and FGL1 (C). HAMP (D) and ID1 (E) expression in Hep3B cells and mouse primary hepatocytes treated for 6h with BMP6 and either Fc, full length FGL1 and the N-terminal or globular domain of FGL1. Hepatic Hamp RNA expression (F), serum hepcidin concentration (G) and liver Id1 (H) and Smad7 (I) mRNA expression in mice treated for 6 hours with saline, Fc or recombinant FGL1 (10 mg/kg) (n=5). Data shown are means ± s.e.m of three independent experiments (A- E) or treated mice and were compared for each condition to untreated cells or control mice by Student t-test. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05.

Article Snippet: Mouse FGL1 cDNA sequences (full length, N-terminal domain, globular domain) were cloned into pFUSEN-hG2Fc plasmid (InvivoGen) with the following modifications: vector signal sequence (from Interleukin-2) was used instead of the native, followed by the Fc fragment of human IgG2.

Techniques: Expressing, Recombinant, RNA Expression, Concentration Assay, Saline, Control

Journal: bioRxiv

Article Title: The hepatokine FGL1 regulates hepcidin and iron metabolism during the recovery from hemorrhage-induced anemia in mice

doi: 10.1101/2023.04.06.535920

Figure Lengend Snippet: Time course of Fgl1 mRNA expression in the liver (A) and the bone marrow (B) 1 to 6 days after phlebotomy in WT and Erfe-/- mice (n=5-8). (C) Hamp and Fgl1 mRNA expression in the liver of 7 week-old mice at t=0 to 20h after a single intra-peritoneal injection of EPO (200u) (n=5). (D) Fgl1 mRNA expression in liver of 8 week-old WT, Th3/+ and Erfe-/-; Th3/+ mice (n=7-9). (F) Fold change of Fgl1 mRNA expression in mouse primary hepatocytes in serum free or serum-containing (5% FBS) media and incubated for 15 hours in low oxygen condition (2%) or in presence of prolyl hydroxylases inhibitor DMOG compared to untreated cells (dashed line). Relative Fgl1 mRNA expression in the liver of Vhl -deficient mice (G), and in the liver of mice treated with prolyl hydroxylase inhibitor vadadustat (H). Data shown are means ± s.e.m and were compared for each time point to values for control WT at t = 0 by Two-way ANOVA (A, B, C) or to WT or vehicle treated mice by Student t-test (D, F, G). Data shown for experiment in primary hepatocytes are means of three independent experiments and were compared to control cells by Student t-test (E). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05.

Article Snippet: Mouse FGL1 cDNA sequences (full length, N-terminal domain, globular domain) were cloned into pFUSEN-hG2Fc plasmid (InvivoGen) with the following modifications: vector signal sequence (from Interleukin-2) was used instead of the native, followed by the Fc fragment of human IgG2.

Techniques: Expressing, Injection, Incubation, Control

Journal: bioRxiv

Article Title: The hepatokine FGL1 regulates hepcidin and iron metabolism during the recovery from hemorrhage-induced anemia in mice

doi: 10.1101/2023.04.06.535920

Figure Lengend Snippet: Fgl1 mRNA expression in the liver of male and female WT mice 36 hours after bleeding compared to control mice (A). Red blood cell count (RBC) (B) and hemoglobin (Hb) (C), marrow (D) and spleen (E) Erfe mRNA expression, liver Hamp (F), Id1 (G), Smad7 (H) mRNA expression in WT and Fgl1-/- mice at t=0 or 36 hours after phlebotomy. Data shown are means ± s.e.m (n= 5-11) and were compared between each group by Two-way ANOVA and corrected for multiple comparisons by Holm-Šidák method. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05.

Article Snippet: Mouse FGL1 cDNA sequences (full length, N-terminal domain, globular domain) were cloned into pFUSEN-hG2Fc plasmid (InvivoGen) with the following modifications: vector signal sequence (from Interleukin-2) was used instead of the native, followed by the Fc fragment of human IgG2.

Techniques: Expressing, Control, Cell Counting

Journal: bioRxiv

Article Title: The hepatokine FGL1 regulates hepcidin and iron metabolism during the recovery from hemorrhage-induced anemia in mice

doi: 10.1101/2023.04.06.535920

Figure Lengend Snippet: Relative expression of Hamp (A), Id1 (B), Smad7 (C) mRNA expression in mouse primary hepatocytes treated with BMP ligand (10ng/ml) and human Fc IgG2 (10µg/ml) or Fc-FGL1 (10µg/ml) for 6 hours. Data shown are means ± s.e.m of three independent experiments and were compared for each BMP between Fc or FGL1 treated cells and control cells by Two-way ANOVA. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05. (D) Western blotting of Hep3B cells treated for 6 hours with BMP6 (10 ng/ml), ERFE (1µg/ml), or FGL1 (10µg/ml) for P-SMAD5, SMAD5 and GAPDH. (E) Western blotting of pull down assay of BMP6 and FGL1 (FL, glob, Nter) for human Fc IgG2 and BMP6.

Article Snippet: Mouse FGL1 cDNA sequences (full length, N-terminal domain, globular domain) were cloned into pFUSEN-hG2Fc plasmid (InvivoGen) with the following modifications: vector signal sequence (from Interleukin-2) was used instead of the native, followed by the Fc fragment of human IgG2.

Techniques: Expressing, Control, Western Blot, Pull Down Assay