Journal: Methods and Protocols

Article Title: Analysis of GPI-Anchored Receptor Distribution and Dynamics in Live Cells by Tag-Mediated Enzymatic Labeling and FRET

doi: 10.3390/mps3020033

Figure Lengend Snippet: Representative kinetics of FRET (dimerization of T-cadherin) following the administration of 10 ug/mL adiponectin in HEK293 living cells. ( a ) The change in the normalized FRET compared to the baseline according to confocal laser scanning microscopy. Each point corresponds to the average NFRET value of a single microscopic image (a few thousand nonzero points). ( b ) The change in the compensated FRET signal measured by flow cytometry. Each point corresponds to the median fluorescence intensity in the 670/30 nm channel after 561 nm excitation of a few thousands living cells gated on the basis of forward and side light scattering. The solid lines represent the fitting of the data by exponential curves.

Article Snippet: After 40–90 s, we added 10 μg/mL recombinant high-molecular-weight-rich human adiponectin (BioVendor, #RD172023100-C).

Techniques: Confocal Laser Scanning Microscopy, Flow Cytometry, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Association between clinical characteristics and serum adiponectin levels in NPC patients.

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

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: The association of serum adiponectin levels with survival in NPC patients. Kaplan–Meier curves were used to illustrate several patients. ( A ) Overall survival and ( B ) metastasis-free survival were analyzed according to serum adiponectin levels based on a cutoff of 8.00 μg/mL in NPC patients ( n = 106). p -values were determined by the log-rank test.

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

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Univariable and multivariable Cox regression analyses of potential factors for the metastasis-free survival in NPC patients a .

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

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Adiponectin suppresses NPC cells invasion and migration. ( A ) Wound healing assay of S18 and 5-8F cells pretreated with adiponectin (0, 10, 20, or 40 μg/mL) for 24 h. Scale bar: 200 μm. Graphs show the relative wound closure. ( B ) Invasion of S18 and 5-8F cells were evaluated by Transwell invasion assay covered with Matrilgel after treatment with various dosing of adiponectin for 24 h. Scale bar: 200 μm. Graphs show the relative number of invasion cells. ( C ) Migration of S18 and 5-8F cells were evaluated by Transwell migration assay after treatment with various doses of adiponectin for 24 h. Scale bar: 200 μm. Graphs show the relative number of migration cells. ( D ) Western blot analysis of E-cadherin, N-cadherin, Slug, Snail, Vimentin, Claudin-1, MMP-2, and MMP-9 in S18 and 5-8F cells after treatment with 40 μg/mL adiponectin for 48 h. 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 adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Migration, Wound Healing Assay, Transwell Invasion Assay, Transwell Migration Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Adiponectin blocks leptin, IL-6, or TNF-α-induced NPC cell invasion and migration. Transwell migration ( A ) and invasion ( B ) assays in S18 and 5-8F cells pretreated with 40 μg/mL adiponectin for 2 h before treatment with 50 ng/mL leptin. Scale bar: 200 μm. Graphs show the relative number of cells. Transwell migration assay in S18 and 5-8F cells pretreated with 40 μg/mL adiponectin for 2 h before treatment with 50 ng/mL TNF-α ( C ) or 50 ng/mL IL-6 ( D ). Scale bar: 200 μm. Graphs show the relative number of migration cells. ( E – G ) Western blot analysis of E-cadherin, N-cadherin, MMP-2, and Vimentin in 5-8F cells pretreated with 40 μg/mL adiponectin in the presence or absence of leptin (50 ng/mL), TNF-α (50 ng/mL), or IL-6 (50 ng/mL) for 48 h. 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 adiponectin was dissolved in deionized water to prepare a working stock solution of approximately 0.5 mg/mL (BioVendor, Brno, Czech Republic).

Techniques: Migration, Transwell Migration Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: Adiponectin inhibits NF-κB and STAT3 activation in NPC cells. Luciferase reporter assay of NF-κB ( A ) and STAT3 ( B ) in S18 and 5-8F cells after treatment with 40 μg/mL adiponectin for 3 h. ( C ) Western blot analysis of p-IκBα, cyto-p65, and nuc-p65 in S18 and 5-8F cells after treatment with various doses of adiponectinfor 6 h. ( D ) Western blot analysis of p-STAT3, cyto-STAT3, and nuc-STAT3 in S18 and 5-8F cells after treatment with various doses of adiponectin for 6 h. Results are presented as mean ± SD of three independent experiments performed in triplicate. ** p < 0.01, *** p < 0.001.

Article Snippet: Recombination human 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, Luciferase, Reporter Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Adiponectin Suppresses Metastasis of Nasopharyngeal Carcinoma through Blocking the Activation of NF-κB and STAT3 Signaling

doi: 10.3390/ijms232112729

Figure Lengend Snippet: AdipoR1 mediates the metastasis inhibitory effect of adiponectin in NPC cells. ( A ) The mRNA expression of AdipoR1 and AdipoR2 was measured with qRT-PCR in S18 and 5-8F cells after transfected with AdipoR1 or AdipoR2 siRNA. ( B ) Transwell migration assay in S18 and 5-8F cells treated with 40 μg/mL adiponectin for 24 h after transfected with AdipoR1 or AdipoR2 siRNA. Scale bar: 200 μm. Graphs show the relative number of migration cells. ( C ) Western blot analysis of E-cadherin, N-cadherin, MMP-9, and MMP-2 in S18 and 5-8F cells treated with 40 μg/mL adiponectin for 48 h after transfected with AdipoR1 siRNA. Results are presented as mean ± SD of three independent experiments performed in triplicate. *** p < 0.001, # p < 0.0001.

Article Snippet: Recombination human 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, Transfection, Transwell Migration Assay, Migration, Western Blot