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| Status |
Public on Feb 05, 2014 |
| Title |
A module-based translational strategy shows a central role for S100A4 in allergy |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
The involvement of thousands of genes complicates the identification of clinically relevant candidate genes in common diseases. We hypothesized that genes co-regulated with a key gene in allergy, IL13, would form a module that could help to discover novel candidate genes. We identified a Th2 cell module by siRNA mediated knock down of 25 putative IL13-regulating transcription factors (TFs) followed by expression profiling.
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| Overall design |
Human CD4+ T cells (hBP CD4+ T cells, 2W-200, Lonza, Vallensbak Strand, Denkmark) were nucleofected either with nucleofection buffer, 1 µM human on target plus SMART pool siRNA against ELK1, GATA3, NFATC3, MAF, NFKB1, JUN, STAT3 (Dharmacon, Lafayette, CO) or non-targeting siRNA using the AMAXA nucleofection program U-014. Six hours after then nucleofection cells were washed, activated and polarized towards Th2. The CD4+ cells were activated with plate bound anti-CD3 (500ng / ml for coating of the plate), with soluble anti-CD28 (500ng / ml) and with IL-2 (17ng /ml, all purchased from R&D). Th2 polarization was induced with anti-IL-12 (5µg/ml) and IL-4 (10ng / ml). For RT-PCR and microarray analyisis the cells were harvested 12 hours of polarization and total RNA was extracted.
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| Contributor(s) |
Bruhn S, Fang Y, Barrenäs F, Gustafsson M, Wang H, Zhao Y, Krönke A, Sönnichse B, Dulyaninova N, Bresnick A, Klingelhöfer J, Nestor C, Xiang Z, Benson M |
| Citation(s) |
24401939 |
| Submission date |
Apr 23, 2013 |
| Last update date |
Oct 11, 2016 |
| Contact name |
Sören Bruhn |
| E-mail(s) |
soren.bruhn@liu.se
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| Organization name |
Linköping University
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| Street address |
Ryddalsgatan 8a
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| City |
Linköping |
| ZIP/Postal code |
582 48 |
| Country |
Sweden |
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| Platforms (1) |
| GPL14550 |
Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Probe Name Version) |
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| Samples (36)
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| Relations |
| BioProject |
PRJNA198904 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE46333_RAW.tar |
111.0 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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