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Status |
Public on Sep 23, 2019 |
Title |
DBA/2J strain CD4 splenocyte at T1, biological rep3 |
Sample type |
RNA |
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Source name |
DBA/2J strain_CD4 splenocyte at T1
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Organism |
Mus musculus |
Characteristics |
strain: DBA/2J gender: female age: 8 weeks tissue: CD4+ splenocytes time point: 4hr after PMA/I stimulation (T1)
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Treatment protocol |
These cells were divided into non-stimulated or phorbol 12-myristate 13-acetate (10 ng ml-1), Boeringer Mannheim) and ionomycin (1uM, Sigma-Aldrich, Missouri)--stimulated (PMA/I) groups and cultured at 37◦C and 5% CO2 for 4 hours.
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Growth protocol |
Splenocytes where extracted and sorted using AutoMACS (Miltenyi Biotech) CD4 Beads (LT34 Miltenyi Biotech) and maintained in MLC supplemented with 10% fetal calf serum (CSL, Parkville, Victoria, Australia) 4 mM L--glutamine, 1 mM sodium pyruvate, 10 mM HEPES, 50 uM beta-mercaptoethanol, penicillin, streptomycin and neomycin.
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Extracted molecule |
total RNA |
Extraction protocol |
Trizol extraction of total RNA was performed followed by clean ups with RNeasy Mini Kits (Qiagen) according to the manufacturer's instructions.
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Label |
biotin
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Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from total RNA. Labelling performed by The Ramaciotti Centre at The University of New South Wales (Sydney, Australia).
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Hybridization protocol |
Standard Affymetrix protocols followed. Hybridisation performed by The Ramaciotti Centre at The University of New South Wales (Sydney, Australia).
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Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 high resolution scanner. Scanning performed by The Ramaciotti Centre at The University of New South Wales (Sydney, Australia).
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Description |
Gene expression data from DBA/2J strain stimulated (PMA/I,4hr) CD4 splenocytes HF_D3S
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Data processing |
The data was RMA normalised with Affymetrix Powertools.
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Submission date |
Mar 15, 2012 |
Last update date |
Sep 23, 2019 |
Contact name |
Hugh John French |
E-mail(s) |
hughjohnfrench@gmail.com
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Organization name |
UNSW
|
Street address |
UNSW
|
City |
Sydney |
State/province |
New South Wales |
ZIP/Postal code |
2036 |
Country |
Australia |
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Platform ID |
GPL6246 |
Series (1) |
GSE36551 |
Influence of genetic variation on inducible gene expression in murine CD4+ splenocytes. |
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