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High throughput screening of lipopolysaccharide treatment in nasal fibroblast

ABSTRACT: To evaluate effect of lipopolysaccharide in nasal fibroblast, we have employed whole genome microarray expression profiling as a discovery platform. Nasal fibroblasts were exposed to LPS (10 μg/mL) for 12 h. Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA). For control and test RNAs, synthesis of target cRNA probes and hybridization were performed using the Low RNA Input Linear Amplification kit (Agilent Technology, Santa Clara, CA). Hybridized images were scanned using a DNA microarray scanner and quantified using Feature Extraction Software (Agilent). All data normalization and selection of fold-change of the genes were performed using GeneSpringGX 7.3 (Agilent).

ORGANISM(S): Homo sapiens

PROVIDER: GSE52505 | GEO | 2015/12/31

SECONDARY ACCESSION(S): PRJNA229132

REPOSITORIES: GEO

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High throughput screening of lipopolysaccharide treatment in nasal fibroblast

Project description:To evaluate effect of lipopolysaccharide in nasal fibroblast, we have employed whole genome microarray expression profiling as a discovery platform. Nasal fibroblasts were exposed to LPS (10 Î¼g/mL) for 12 h. Total RNA was isolated using Trizol reagent (Invitrogen, Carlsbad, CA). For control and test RNAs, synthesis of target cRNA probes and hybridization were performed using the Low RNA Input Linear Amplification kit (Agilent Technology, Santa Clara, CA). Hybridized images were scanned using a DNA microarray scanner and quantified using Feature Extraction Software (Agilent). All data normalization and selection of fold-change of the genes were performed using GeneSpringGX 7.3 (Agilent). Nasal fibroblasts were exposed to LPS (10 Î¼g/mL) for 12 h. After exposure for 12hr with LPS, the untreated nasal fibroblasts and LPS-treated nasal fibroblasts were harvested and measured. Double independent experiments were performed at each time (0 or 12 hours) using different nasal fibroblasts for each experiment. So these data are duplicated.

2015-12-31 | E-GEOD-52505 | biostudies-arrayexpress

Expression data from CRS with nasal polyps (CRSwNP) and normal control

Project description:Chronic rhinosinusitis (CRS) is a prevalent chronic rhinologic disease, characterized by persistent symptomatic inflammation of the nasal mucosa and paranasal sinuses. Sinonasal tissues were obtained during sinus surgeries performed at Northwestern University. For the microarray, NP tissues and uncinate tissues from normal healthy controls were divided into elderly (≥65 years old, each n=3-4), or non-elderly (18-49 years old, each n=3-4) according to age. This study protocol was approved by the independent ethics committees or institutional review boards (IRB) at Northwestern University. Total RNA was isolated from NP tissues and control uncinate tissues. RNA concentration was calculated, and the highest quality RNA used for microarray. The quality of total RNA was evaluated by Bioanalyzer (Agilent Technologies, Santa Clara, CA). The samples were at a concentration ≥125ng/μl and a A260/A280 ratio that was ≥1.80 and a RIN value that was ≥7.0 were considered acceptable. The highest quality RNA was used to generate labeled cRNA and hybridized to Affymetrix Human Gene 2.1ST array following the manufacturer’s protocol (Affymetrix Inc., Santa Clara, CA). Chip processing and image capturing were performed using Affymetrix AGCC software. We investigated age- and disease-related differential gene expression in chronic rhinosinusitis with nasal polyps.

2022-01-27 | GSE194282 | GEO

Mice liver and urine LC-MS data

Project description:Mouse livers were analyzed by lipidomics and urine by metabolomics.An Agilent Ultra-Performance Liquid Chromatography/Electrospray Ion Quadrupole Time-of-Flight-Mass Spectrometer (UPLC-ESI-QTOFMS) (Agilent, Santa Clara, CA) was utilized for lipid and other metabolites proofing in this work.

2023-06-07 | PXD041465 | Pride

Modulation of microglia-mediated amyloid-beta clearance with nanoformulated polyphenol through the SIRT1-LDLR pathway

Project description:Light-controlled in situ synthesis of DNA microarrays (Affymetrix GeneChip® Mouse Genome 430 2.0) were performed to determine the altered gene expression. Affymetrix algorithm and multiple analysis comparison software were used for assessing gene expression differences, and mRNAs that increased or decreased in the brains of NP(α-M)-treated mice relative to that in the brains of Blank-NP-treated mice were identified. For the assay, total RNA was extracted using Takara RNAiso Plus Kit (Cat#9109, Takara, DaLian, LiaoNing, CN) following the manufacturer’s instructions and checked for a RIN number to inspect RNA integrity by an Agilent Bioanalyzer 2100 (Agilent technologies, Santa Clara, CA, US). Qualified total RNA was further purified by RNeasy mini kit (Cat#74106, QIAGEN, GmBH, Germany) and RNase-Free DNase Set (Cat#79254, QIAGEN, GmBH, Germany). Array hybridization and wash was performed using GeneChip® Hybridization, Wash and Stain Kit (Cat#900720, Affymetrix, Santa Clara, CA, US) in Hybridization Oven 645 (Cat#00-0331-220V, Affymetrix, Santa Clara, CA, US) and Fluidics Station 450 (Cat#00-0079, Affymetrix, Santa Clara, CA, US) followed the manufacturer’s instructions. Slides were scanned by GeneChip® Scanner 3000 (Cat#00-00212, Affymetrix, Santa Clara, CA, US) and Command Console Software 4.0 (Affymetrix, Santa Clara, CA, US) with default settings. Raw data were normalized by MAS 5.0 algorithm, Gene Spring Software12.6.1 (Agilent technologies, Santa Clara, CA, US).

2018-09-14 | GSE119904 | GEO

Microarray analysis of lung SPC/GFP+ EpCAM+ cells

Project description:We extracted RNA of sorted lung SPC/GFP+ EpCAM+ cells and performed microarray analyses. Total RNA was extracted from sorted Ep-CAMhigh/GFPhigh cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. The RNA integrity was examined on an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA). Biotinylated ss-cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA using the GeneChip WT PLUS Reagent Kit User Manual (Affymetrix/Thermo Fisher Scientific). Fragmented and labeled ss-cDNA were hybridized on a GeneChip Clariom S Array (Affymetrix/Thermo Fisher Scientific) (n =3/group). We used microarrays to detail the global gene expression of SPC/GFP+ EpCAM+ cells from the control mice(G29-5_(Clariom_S_Mouse)) and the 3 months smoke mice(G29-6_(Clariom_S_Mouse)), and identified distinct classes of up-regulated genes during this process.

2019-04-26 | GSE130331 | GEO

microarray analysis of lung SPC/GFP+ EpCAM+ cells from different smoke exposure mice

Project description:We extracted RNA of sorted lung SPC/GFP+ EpCAM+ cells and performed microarray analyses. Total RNA was extracted from sorted Ep-CAMhigh/GFPhigh cells using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. The RNA integrity was examined on an Agilent 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA). Biotinylated ss-cDNA were prepared according to the standard Affymetrix protocol from 100 ng total RNA using the GeneChip WT PLUS Reagent Kit User Manual (Affymetrix/Thermo Fisher Scientific). Fragmented and labeled ss-cDNA were hybridized on a GeneChip Clariom S Array (Affymetrix/Thermo Fisher Scientific) (n =3/group). We used microarrays to detail the global gene expression of SPC/GFP+ EpCAM+ cells from the control mice(G37-1_(Clariom_S_Mouse)), 3 week continuous smoke mice (G37-2_(Clariom_S_Mouse)) and 3 weeks intermittent smoke mice(G37-3_(Clariom_S_Mouse)), and identified distinct classes of up-regulated genes during this process.

2020-12-10 | GSE162919 | GEO

Integrative array-based approach identifies MZB1 as a frequently methylated putative tumor-suppressor in hepatocellular carcinoma (MeDIP)

Project description:The analysis of DNA methylation profiles was performed by methylated DNA immunoprecipitation (MeDIP) using anti-5-methylcytidine antibody (Eurogentec, Seraing, Belgium) followed by the Human 244K Agilent CpG island microarray (Agilent, Santa Clara, CA)

2012-03-31 | GSE35312 | GEO

Expression of cyclophilin B is associated with malignant progression and regulation of genes

Project description:Microarray was conducted on Illumina Human Ref-6 Version 2 Expression Chip (Illumina, San Diego, CA). Three independent cultures were used for RNA isolation with RNeasy plus mini kit (Qiagen, Valencia, CA). RNA quality was checked by Agilent Bioanalyzer (Agilent, Santa Clara, CA). An Ambion labeling kit was used for labeling cDNA followed by hybridization to Illumina chips. ChIP scan data was extracted by Illumina Beadstudio and subsequently analyzed using Bioconductor lumi package.

2009-04-03 | GSE15505 | GEO

RNA sequencing in human iPSCs derived from bipolar patients to identify important therapeutic molecular targets of Valproate(VPA)

Project description:Valproate(VPA) has been used in the treatment of bipolar disorder since the 1990s. However, the therapeutic targetsof VPA have remained elusive. Here we used RNA sequencing in human iPSCs derived from bipolar patients to further identify important molecular targets. Human iPSCs were homogenized and total RNA was isolated using the RNeasy Plus Micro Kit (Qiagen, Hilden, Germany). RNA quantity and quality were assessed using fluorometry (Qubit RNA Broad Range Assay Kit and Fluorometer; Invitrogen, Carlsbad, CA) and chromatography (Bioanalyzer and RNA 6000 Nano Kit; Agilent, Santa Clara, CA), respectively. Libraries were prepared using TruSeq Stranded mRNA (PolyA+) kit (Illumina, San Diego, CA) and sequenced by Illumina NextSeq 500. The read length was 75bp with 30-40M reads per sample. FastQC (v0.11.3) was performed to assess data quality. TopHat2 (v2.0.9) aligned the reads to the mouse reference genome (Mus musculus UCSC mm10) and to the Ensembl human reference genome (GRCh38.p13) using default parameters. Alignments were then converted to expression count data using HTseq (v0.6.1) with default union mode.

2020-12-31 | GSE160761 | GEO

Combination of ABT-869 with SAHA on AML with FLT3-ITD

Project description:For the microarray experiments, MV4-11 and MOLM-14 cells were treated with DMSO control, ABT-869 3 nM, SAHA 6 uM and combination therapy for 24 hours. Cells were then washed in PBS and high-quality total RNA was extracted RNeasy Midi Kit, according to the manufacturer’s instruction (Qiagen, Valencia, USA). RNA quantity, quality, and purity were assessed with the use of the RNA 6000 Nano assay on the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara CA, USA). Gene expression profiling was performed using Affymetric U133plus2.0 gene chip (Affymetrix, Santa Clara, CA, USA) according to the manufacturer’s protocol.

2011-05-31 | GSE26114 | GEO

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