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Series GSE81527 Query DataSets for GSE81527
Status Public on May 18, 2016
Title IRAK4_hPBMC_stimulus_compound_exp3
Organism Homo sapiens
Experiment type Expression profiling by array
Summary The goal of this experiment is to characterize the ability of IRAK4 kinase inhibitors to block TLR7 or TLR9 induced gene transcription in human PBMCs. PBMCs from two donors were pretreated with compound for 30 minutes and stimulated with gardiquimod (TLR7 ligand) or ODN 2216 (TLR9 ligand) for 5 hrs. Compound treated gene expression profiles wil be compared to untreated.
 
Overall design PBMCs from 2 independent donors were isolated, pretreated with 5 μM BMS-986126 (BMT-087965) or DMSO (vehicle) for 30 minutes at 37°C and then stimulated for 5 hours with 5 μg/ml gardiquimod. Total RNA was isolated from the cells with Qiagen RNeasy Mini Kit (Cat. # 74104). Total RNA was then treated with DNase I and cleaned up with Qiagen RNeasy MiniElute Cleanup Kit (Cat. # 74204). The RNA concentrations were determined by NanoDrop and RNA quality was evaluated using Experion (Bio-Rad). All target labeling reagents were purchased from Affymetrix (West Sacramento CA). Double-stranded cDNAs were synthesized from 1ug of total RNA through reverse transcription with an oligo-dT primer containing the T7 RNA polymerase promoter and double strand conversion using the cDNA Synthesis System. Biotin-labeled cRNA was generated from the cDNA and used to probe Human Genome U219 array, comprising of 49,293 probe sets and representing more than 20,000 human genes, using the Gene Titan MC instrument (Affymetrix, West Sacramento, CA). All cDNA and cRNA target preparation steps were processed on a Caliper GeneChip Array Station from Affymetrix. Array hybridization, washing and scanning were performed according to the manufacturer’s recommendations.
 
Contributor(s) Julie C
Citation(s) 28003376
Submission date May 17, 2016
Last update date Mar 21, 2019
Contact name Hanjoong Jo
E-mail(s) hanjoong.jo@bme.gatech.edu
Phone (404) 712-9654
Organization name Georgia Tech and Emory Univ
Department Coulter Department of Biomedical Engineering
Street address 1760 Haygood Drive, Health Sciences Research Bldg E170
City Atlanta
State/province GA
ZIP/Postal code 30322
Country USA
 
Platforms (1)
GPL13667 [HG-U219] Affymetrix Human Genome U219 Array
Samples (18)
GSM2155122 IRAK4_EPH06P1_hPBMC_GARD_vehicle_D122_2C
GSM2155123 IRAK4_EPH03P1_hPBMC_GARD_BMT087965_D122_5C
GSM2155124 IRAK4_EPG05P1_hPBMC_unstim_vehicle_D140_1E
Relations
BioProject PRJNA321911

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE81527_RAW.tar 62.6 Mb (http)(custom) TAR (of CEL)
Processed data included within Sample table

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