Project description:Non-coding RNA profiling by microarray in an Affymetrix® miRNA Array, version 4.1 (Santa Clara, CA, USA), commercially available by Thermo Fisher Scientific (Waltham, MA) We aimed to profile the expression of the entire population of annotated small non-coding RNAs in the Temporal Cortex (TC) of Alzheimer diseased (AD) individuals compared to age-matched controls.
Project description:To investigate the molecular pathological mechanisms of irritable bowel syndrome with diarrhea (IBS-D) and elucidate the effects of acupuncture on IBS-D colonic mucosa protein abundance in rats, a label-free high-throughput liquid chromatography-tandem mass spectrometry (LC-MS)-based proteomics analysis was used to survey the global changes of colonic mucosa proteins between different groups. A Nano flow Ultimate 3000 HPLC (Dionex Corp, Sunnyvale, CA) coupled online to a Q-Exactive Plus mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) was used in this project.
Project description:Tandem Mass Tag (TMT) technology is an in vitro peptide labeling technique developed by Thermo Fisher Scientific, a peptide labeling technology developed in vitro. The technology uses six (TMTsixplex™ Isobaric Label Reagent Set) or 10 (TMT10plex™ Isobaric Label Reagent Set) isotope labels by labeling peptide specific amino acid sites. In TMT experiments, the relative and absolute amounts of proteins in up to 10 different samples can be compared flexibly by tandem mass spectrometry analysis. In TMT experiments, whole proteins from tissue cells are obtained by protein extraction, and the whole proteins are enzymatically cleaved into peptides by trypsin, and the TMT reagents are used to label all the obtained peptides. The TMT reagent consists of three main components: a reporter group, an equilibrium group and a peptide reaction group.
Project description:To explore the cellular pathways and the molecular functions affected by a novel biogenic amine, (3-HKA), we performed a label free quantitative (LFQ) proteomic analysis of mouse lymphatic endothelial cells (LEC), and primary dendritic cells, treated with IFNgammaplus or minus 3-HKA. Biological triplicates, of total cell lysates, were fractionated by one-dimensional gel electrophoresis (1DEF) and the “in gel” tryptic derived peptides analyzed by nano-LC-ESI-MS/MS on a Q Exactive HF quadrupole orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). LFQ analysis highlighted that 3-HKA downregulated many of the inflammatory pathways, more notably JAK/STAT1 and NFkB, associated with IFNgamma activation.
Project description:Affymetrix Human Gene 2.0 ST microarray (ThermoFisher Scientific, Waltham, MA, USA) was used to select differentially expressed genes.
Project description:ARPE-19 RPE cells obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) were cultured in DMEM medium with 10% FBS (Thermo Fisher Scientific, Waltham, USA) at the incubator with 37 °C, 5% CO2 and 100% humidity. Final concentration of 15 μM curcumin (C7727, Sigma-Aldrich, St. Louis, USA) and 100 μM CoCl4 (10007216, Sinopharm Chemical Reagent, Shanghai, China) were added within serum-free medium for 24 h and 4 h respectively. RNA-seq was performed to investigate the transcriptome alteration.
Project description:Purpose: The purpose of this study is to measure the changes in liver transcriptome in response to short-term fasting between 7 and 13 h where the rats were dosed with 2 ml/kg of saline vehicle at 0 h Methods: Total RNA was isolated from the liver, using TRIzol Reagent (Thermo Fisher Scientific, Waltham, MA) and the direct-zol RNA Mini Prep kit (Zymo Research, Irvine, CA). The isolated RNA samples were then submitted to the Vanderbilt University Medical Center VANTAGE Core (Nashville, TN) for RNA quality determination and sequencing. Total RNA quality was assessed using a 2100 Bioanalyzer (Agilent, Santa Clara, CA). At least 200 ng of DNase-treated total RNA with high RNA integrity was used to generate poly-A-enriched mRNA libraries, using KAPA Stranded mRNA sample kits with indexed adaptors (Roche, Indianapolis, IN). Library quality was assessed using the 2100 Bioanalyzer (Agilent), and libraries were quantitated using KAPA library Quantification kits (Roche). Pooled libraries were subjected to 75-bp paired-end sequencing according to the manufacturer’s protocol (Illumina HiSeq3000, San Diego, CA). Results: No genes were were found to be differentially expressed with a false discovery rate less than 0.1 Conclusions: There were no significant changes in liver gene expression between 7 and 13 h of fasting
