|
| Status |
Public on Nov 23, 2012 |
| Title |
Human_ThGARP_T360min |
| Sample type |
RNA |
| |
|
| Source name |
human peripheral blood of healthy donor
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: sorted human Teff GARP-overexpressed cells (CD4+CD25-) stimulated by anti-CD3/anti-CD28 Dynal beads/IL2
|
| Treatment protocol |
Human Tregs or Teffs were mixed with anti-CD3/anti-CD28-coated Dynabeads (Invitrogen) at a ratio of 1:1 and with 100 U/ml of IL2 (Proleukin, Novartis) and were distributed with 4x106 cells into 1.5 ml microtubes (Eppendorf) for each time point. Before the cells were cultured at 37°C, they were first stored at 4°C to allow cells and beads to settle properly. Samples were taken every 20 min during the first six hours following stimulation, except for TeffGARP and TeffGFP cells, which were measured at time points 0, 100 and 360 min. The cells were centrifuged, and the cell pellets were lysed in RLT buffer (Qiagen) and immediately stored at -70°C.
|
| Growth protocol |
taken from donors, sorted, expanded ex vivo and stimulated
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 microg total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 microg of cRNA were hybridized for 16 hr at conditions recommended by the manufacturer. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GCS 3000.
|
| Description |
transcriptomic data to construct a dynamic functional network of human regulatory T cell genes
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 150.
|
| |
|
| Submission date |
Apr 29, 2008 |
| Last update date |
Oct 23, 2013 |
| Contact name |
Feng Q. Hefeng |
| E-mail(s) |
feng.hefeng@lih.lu
|
| Organization name |
Luxembourg Institute of Health
|
| Department |
Department of Infection and Immunity
|
| Street address |
29, rue Henri Koch
|
| City |
Esch-sur-Alzette |
| ZIP/Postal code |
4354 |
| Country |
Luxembourg |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE11292 |
High-time-resolution dynamic analysis of human regulatory T cell (Treg) / CD4+ T-effector cell (Teff) activation |
|
