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: eLife

Article Title: An adipokine feedback regulating diurnal food intake rhythms in mice

doi: 10.7554/eLife.55388

Figure Lengend Snippet: ( A ) Representative normalized Bmal1-luc luminescence rhythms of dexamethasone (DEX) synchronized hypothalamus-derived mHypo-N44 ( N44 ) cells. Inset depicts raw luminescence data of the same set. ( B ) Normalized Bmal1-luc luminescence rhythms of N44 cells after treatment with ADIPOQ (Adn; red and blue) peptide or PBS (black) at the depicted time points after synchronization. Shown are averages ± SEM of 3 traces each. ( C ) Phase response curve for ADIPOQ-mediated resetting of N44/ Bmal1-luc cells (n = 3 per time point; treatment time given in degrees with 90 °=maximal luminescence and 270 °=minimal luminescence – see ( A ) for reference). ( D ) Normalized PER2::LUC luminescence rhythms of primary hypothalamic neurons after treatment with ADIPOQ (Adn; red and blue) peptide at the depicted time points after synchronization. Shown are averages ± SEM of 3 traces each. Inset shows quantification of phase shifts (p<0.0001; unpaired Student’s t-test). ( E ) Normalized PER2::LUC luminescence rhythms of organotypic MBH slices after treatment with ADIPOQ (Adn; red and blue) peptide at the depicted time points. Shown are averages ± SEM of 3 traces each. ( F ) Phase response curve for ADIPOQ-mediated resetting of PER2::LUC MBH slices (treatment time given in degrees with 90 °=maximal luminescence and 270 °=minimal luminescence – see ( E ) for reference. Figure 5—source data 1. Raw data of experiments shown in .

Article Snippet: Forskolin (Fors), dexamethasone (Dex), globular adiponectin (Adn), and AdipoRon were purchased from Phoenix Pharmaceuticals (Karlsruhe, DE); leptin and full-length adiponectin were purchased from Enzo Life Science (Loerrach, DE); ghrelin, glucagon, and nesfatin were purchased from Bachem (Bubendorf, CH); resistin was purchased from Prospec (NJ, USA) and visfatin from Aviscera Bioscience (CA, USA); VPR66 was purchased from BioMol (Hamburg, DE).

Techniques: Derivative Assay

Journal: eLife

Article Title: An adipokine feedback regulating diurnal food intake rhythms in mice

doi: 10.7554/eLife.55388

Figure Lengend Snippet: ( A, B ) Normalized Bmal1-luc luminescence rhythms of N44 cells after treatment with dexamethasone (Dex; A ) or forskolin (Fors; B ) or PBS at the depicted time points after synchronization. Shown are averages ± SEM of 3 traces each. Bottom panels: phase response curves for dexamethasone (left) and forskolin (right). Shown are averages of three experiments per time point each with sine wave regressions (treatment time given in degrees with 90 °=maximal luminescence and 270 °=minimal luminescence – see for reference.

Article Snippet: Forskolin (Fors), dexamethasone (Dex), globular adiponectin (Adn), and AdipoRon were purchased from Phoenix Pharmaceuticals (Karlsruhe, DE); leptin and full-length adiponectin were purchased from Enzo Life Science (Loerrach, DE); ghrelin, glucagon, and nesfatin were purchased from Bachem (Bubendorf, CH); resistin was purchased from Prospec (NJ, USA) and visfatin from Aviscera Bioscience (CA, USA); VPR66 was purchased from BioMol (Hamburg, DE).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Interaction of Nerve Growth Factor β with Adiponectin and SPARC Oppositely Modulates its Biological Activity

doi: 10.3390/ijms20071541

Figure Lengend Snippet: Summary of analyte binding to adiponectin.

Article Snippet: Briefly, full length adiponectin, globular adiponectin or SPARC (Sangi, Tokyo, Japan) as a ligand was immobilized onto the carboxymethylated dextran surface of the CM5 sensor chip, respectively.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Interaction of Nerve Growth Factor β with Adiponectin and SPARC Oppositely Modulates its Biological Activity

doi: 10.3390/ijms20071541

Figure Lengend Snippet: Adiponectin suppressed NGFβ-induced neurite outgrowth and cell swelling. ( A , B ) PC12 cells were treated with increasing concentration of NGFβ either in the presence or absence of full-length adiponectin (fADPN, 1 µg/mL) and globular adiponectin (gADPN, 1 µg/mL). Representative results of the cells (arrowhead: neurite) are shown in A and results (percentage of the cells with axon) from five independent experiments are summarized in B. ( C , D ) PC12 cells were treated with NGFβ (1 ng/mL) either in the presence or absence of full length adiponectin and globular adiponectin (0.1 and 1 g/mL). The ratio of the cell with axon ( C ) and the changes in cell body size ( D ) are determined and summarized from three independent experiments. * indicates the statistically significant difference ( p < 0.05) from NGFβ treatment alone (Cont).

Article Snippet: Briefly, full length adiponectin, globular adiponectin or SPARC (Sangi, Tokyo, Japan) as a ligand was immobilized onto the carboxymethylated dextran surface of the CM5 sensor chip, respectively.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Interaction of Nerve Growth Factor β with Adiponectin and SPARC Oppositely Modulates its Biological Activity

doi: 10.3390/ijms20071541

Figure Lengend Snippet: Adiponectin suppressed NGFβ-induced neurite outgrowth independently of its receptor activation. ( A ) Expression of AdipoR1 and AdipoR2 mRNA in the rat skeletal muscle (SM), liver and PC12 cells are shown. ( B ) PC12 cells were treated with full length adiponectin or globular adiponectin and the amounts of phosphorylated and total AMPK were determined. Representative results and the ratio of phosphorylated and total AMPK are shown ( n = 5). ( C – E ) PC12 cells were treated with unrelated (un), AdipoR1, AdipoR2 and R1 plus R2 siRNA and ( C ) mRNA expression of AdipoR1 and AdipoR2 are shown. ( D ) The transfected cells were treated with vehicle (cont.), globular adiponectin (1 µg/mL) and full length adiponectin (1 µg/mL) and the state of AMPK activation are shown ( n = 3). ( E ) The transfected cells were treated with vehicle (cont.), NGFβ (1 ng/mL) or NGFβ plus full length adiponectin (1 µg/mL) and the ratios of the cell with axon are shown ( n = 3). The transfected cells treated with vehicle did not induce any neurite (axon) as shown in A and the ratio calculated was 0 as in B. Thus, bar for control value of each siRNA was not seen. * indicates the statistically significant difference ( p < 0.05) from cont. or NGFβ treatment alone.

Article Snippet: Briefly, full length adiponectin, globular adiponectin or SPARC (Sangi, Tokyo, Japan) as a ligand was immobilized onto the carboxymethylated dextran surface of the CM5 sensor chip, respectively.

Techniques: Activation Assay, Expressing, Transfection, Control