Journal: PLoS ONE

Article Title: The Role of PPARγ in Advanced Glycation End Products-Induced Inflammatory Response in Human Chondrocytes

doi: 10.1371/journal.pone.0125776

Figure Lengend Snippet: In A, chondrocytes were pretreated with anti-RAGE (5 μg/ml) for 12 hours before AGEs (100 μg/ml) stimulation. In B, chondrocytes were pretreated with SB203580, SP600125, PD98059 (10 μM) for 30 minutes prior to AGEs (100 μg/ml) stimulation. The expression of PPARγ was quantified by real-time PCR and western blotting using β-actin as an internal control. In A, densitometric analysis for PPARγ levels corrected to β-actin is shown. All data are expressed as means ± SD and are representative of three independent experiments. *: p< 0 . 05 versus control, #: p< 0 . 05 versus AGEs treatment, &: p> 0 . 05 versus AGEs treatment.

Article Snippet: Rabbit monoclonal antibodies specific for IL-1β, NF-κB p65, PPARγ, TNF-α, IκBα, β-actin and RAGE were purchased from Cell signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Molecular immunology

Article Title: S100A11 regulates nasal epithelial cell remodeling and inflammation in CRSwNPs via the RAGE-mediated AMPK-STAT3 pathway.

doi: 10.1016/j.molimm.2021.09.014

Figure Lengend Snippet: Fig. 5. S100A11 depends on RAGE-mediated can activate AMPK and inhibit STAT3 signaling pathway. A. The phosphorylation levels of AMPK and STAT3 were detected by Western blot after S100A11 was overexpressed. B. The histogram shows the phosphorylation ratio of AMPK and STAT3 (pAMPK/ AMPK and pSTAT3/ STAT3). C. Western blot showed the expression of cleaved-PARP, CCND1, CCNE, Bcl-2, AMPK and STAT3 phosphorylated proteins after treatment of cells with different concentrations of rh-S100A11 protein (0~10000ug/mL) for 24 h. D. Cell immunofluorescence staining detected the expression of STAT3 phosphorylated protein after treatment with 1000ug/mL rh-S100A11 protein for 24 h (bar = 20um). E. The relative mRNA levels of Fas, Stk35, Tsc2 and Socs3 after cells were treated with 1000ug/mL rh-S100A11 protein or transfected for 24 h. F. RAGE protein inhibitors affect the phosphorylation levels of AMPK and STAT3 caused by rhS100A11. The bar graphs show quantification of the results, with each value represents the mean ± SD of three independent experiments. Statistical significance is shown using the Student’s t-test analysis, *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Rabbit polyclonal anti− CCND1 antibody (Cat. No. 60,186-1-Ig, 1:2000 diluted), anti-RAGE antibody (Cat. No. 66,833-1-Ig, 1:2000 diluted) and anti− CCNE antibody (Cat. No. 11554− 1-AP, 1:2000 diluted for WB) were purchased from Proteintech.

Techniques: Phospho-proteomics, Western Blot, Expressing, Immunofluorescence, Staining, Transfection

Journal: Journal of food and drug analysis

Article Title: Attenuation of diabetes-mediated muscle atrophy in rats by fish oil enriched omega-3 polyunsaturated fatty acids supplementation.

doi: 10.38212/2224-6614.3468

Figure Lengend Snippet: Fig. 9. Proposed molecular mechanisms for the benefits of fish oil on muscle atrophy triggered by diabetes. Fish oil improves skeletal muscle health through enhancing myofibril protein synthesis, reducing myofibril protein degradation (UPS and ALS), relieving inflammation, and improving mitochondria function, which may lead to net attenuate muscle atrophy along with improved insulin sensitivity. Abbreviations: AGEs, advanced glycation end products; AMPK, adenosine monophosphate-activated protein kinase; ATG5, autophagy protein 5; FoxO1, forkhead box O 1 tran- scription factor; LC3B II, microtubule-associated protein light chain 3 b II; mTOR, mammalian target of rapamycin; MURF1, muscle RING-finger protein 1; p62, p62/sequestosome 1; p70s6k, p70 ribosomal S6 kinase; PGC-1a, peroxisome proliferator-activated receptor-gamma coactivator 1 a; RAGE, receptor for AGEs; SIRT1, sirtuin1.

Article Snippet: Afterwards, membranes were blocked with 5% skim milk in 1x tris-buffered saline with Tween (TBST) at room temperature for 1 h and then incubated with primary antibodies (1:1000 v/v dilution) for phosphorylated FoxO1a (Ser256), FoxO1a, phosphorylated AKT (Ser473), AKT, phosphorylated NF-kB-p65 (Ser536), NF-kB-p65, Atrogin-1, PGC-1a (Abcam, Cambridge, MA, USA), phosphorylated AMPKa (Thr172), AMPKa (Cell Signaling Technology, Danvers, MA, USA), MURF1, phosphorylated mTOR (Ser2481), mTOR, and ATG5, p62, LC3BI, LC3BII, phosphorylated p70s6k (Ser434), p70s6k, RAGE, SIRT1 and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at 4 C. Following primary antibody incubation, membranes were washed with 1x TBST and incubated with horseradish peroxidase-conjugated secondary antibodies (1:1000 v/v dilution, Cell Signaling Technology) for 1 h at room temperature.

Techniques: