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Status |
Public on Aug 01, 2017 |
Title |
WT FoxP3-transduced T cells_3 |
Sample type |
RNA |
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Source name |
GFP(high) CD4(+) T cells transduced with WT FoxP3/MIGR1 retroviral vector
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Organism |
Mus musculus |
Characteristics |
strain: BALB/cA tissue: lymph nodes and spleen cell type: In vitro activated CD4(+) T cells transduction: WT FoxP3/MIGR1 retroviral vector
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Treatment protocol |
Sorted naïve CD4(+) T cells were stimulated with 0.5 ug/ml anti-CD3e (145-2C11) in the presence of 10 ng/ml recombinant mouse IL-2 and syngeneic T cell-depleted and irradiated (20Gy) spleen cells, and spin-infected with retroviral supernatants 24 hr later.
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Growth protocol |
Retrovirally transduced T cells were cultured in RPMI1640 medium supplemented with 10% FBS, 10 mM HEPES, 1 mM Sodium Pyruvate, 50 uM 2-ME, PN/ST, and 10 ng/ml recombinant mouse IL-2.
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted using Isogen reagent (Nippon Gene) followed by RNeasy mini kit (Qiagen).
|
Label |
biotin
|
Label protocol |
RNA was subjected to biotinylated cDNA synthesis by using GeneChip One-Cycle Target Labeling and Control Reagents.
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Hybridization protocol |
The labeled cDNA was hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 Arrays using Affymetrix standard procedures.
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Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
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Description |
Naïve CD4(+) T cells were transduced with WT FoxP3/MIGR1 retroviral vector and GFP(high) CD4(+) cells were sorted on day 7.
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Data processing |
Expression values for each probe set were calculated with rma method in a GeneSpring GX software package (version 13.0) (Agilent).
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Submission date |
Nov 08, 2016 |
Last update date |
Aug 01, 2017 |
Contact name |
Shohei Hori |
E-mail(s) |
[email protected]
|
Phone |
+81-45-503-7069
|
Organization name |
RIKEN Center for Integrative Medical Sciences
|
Lab |
Laboratory for Immune Homeostasis
|
Street address |
1-7-22 Suehiro-cho, Tsurumi-ku
|
City |
Yokohama |
State/province |
Kanagawa |
ZIP/Postal code |
230-0045 |
Country |
Japan |
|
|
Platform ID |
GPL1261 |
Series (2) |
GSE89645 |
Expression data from mutant FoxP3-transduced CD4 T cells |
GSE89744 |
Analyses of a mutant FoxP3 allele reveal BATF as a critical transcription factor in the differentiation and accumulation of tissue regulatory T cells |
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