|
Status |
Public on Aug 01, 2007 |
Title |
Foxp3 mutant |
Sample type |
RNA |
|
|
Source name |
CD4+ T cells gene-transduced with a Foxp3 mutant defective in binding to AML1.
|
Organism |
Mus musculus |
Characteristics |
T cells were obtained from BALB/c mice and puridied to CD4+ naive T cells (GITRlowCD25-CD4+) and retrovirally gene transduced
|
Treatment protocol |
CD4+ naive T cells were activated and retrovirally gene transducedwith either empty vector (pMCsIg) or AML1-, wild type Foxp3-, or an AML1-non binding mutant Foxp3. Sixty hours after transfection, GFP+ cells were sorted and lysed for RNA samples.
|
Growth protocol |
Naive CD4+ T cells were activated by soluble anti-CD3 Ab [0.5ug/ml], IL-2 [20u/ml] and antigen presenting cells (T cell-depleted, mitomycin-C-treated splenocytes). Twenty-four hours after transfection, retroviral supernatant and polybrene were added and centrifuged for one hour at 3200 rpm. Cells were kept at 37 C, 5% CO2 until they were sorted.
|
Extracted molecule |
total RNA |
Extraction protocol |
We performed RNA extraction using QIAGEN's RNeasy micro columns. according to the manufacteur's protocol.
|
Label |
biotin
|
Label protocol |
One cycle target labelling according to the protocol provided by Affymetrix
|
|
|
Hybridization protocol |
Labelled cRNA (15ug) were hybridized with Mouse Genome 430 2.0 Array for 16 hours, washed, and stained by streptavidin-phycoerthrin.
|
Scan protocol |
The arrays were scanned and aquired data were analyzed by Gene chip operating software (GCOS).
|
Description |
Refer to Ono, M. et al, Nature 2007 (Supplementary Figure S7).
|
Data processing |
Gene chip operating software (GCOS).
|
|
|
Submission date |
Feb 02, 2007 |
Last update date |
Aug 28, 2018 |
Contact name |
Masahiro Ono |
E-mail(s) |
m.ono@ucl.ac.uk
|
Organization name |
Institute of Child Health
|
Department |
Immunobiology Unit
|
Street address |
30 Guilford Street
|
City |
London |
ZIP/Postal code |
WC1N 1EH |
Country |
United Kingdom |
|
|
Platform ID |
GPL1261 |
Series (1) |
GSE6939 |
CD4+ T cells gene-transduced with AML1, wild type Foxp3, and a Foxp3 mutant defective in binding to AML1 |
|
Relations |
Reanalyzed by |
GSE119085 |