|
| Status |
Public on Aug 01, 2017 |
| Title |
R397W FoxP3-transduced T cells_1 |
| Sample type |
RNA |
| |
|
| Source name |
GFP(high) CD4(+) T cells transduced with R397W FoxP3/MIGR1 retroviral vector
|
| Organism |
Mus musculus |
| Characteristics |
strain: BALB/cA
tissue: lymph nodes and spleen
cell type: In vitro activated CD4(+) T cells
transduction: R397W FoxP3/MIGR1 retroviral vector
|
| 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.
|
| 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.
|
| 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.
|
| |
|
| Hybridization protocol |
The labeled cDNA was hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 Arrays using Affymetrix standard procedures.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
|
| Description |
Naïve CD4(+) T cells were transduced with R397W FoxP3/MIGR1 retroviral vector and GFP(high) CD4(+) cells were sorted on day 7.
|
| Data processing |
Expression values for each probe set were calculated with rma method in a GeneSpring GX software package (version 13.0) (Agilent).
|
| |
|
| Submission date |
Nov 08, 2016 |
| Last update date |
Aug 01, 2017 |
| Contact name |
Shohei Hori |
| E-mail(s) |
shohei.hori@riken.jp
|
| 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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