|
| Status |
Public on Jan 17, 2009 |
| Title |
activated CD4+ T cell_plusIL-6-B |
| Sample type |
RNA |
| |
|
| Source name |
CD3/CD28-activated NK1.1-CD4+ T cells/IL-6 treated
|
| Organism |
Mus musculus |
| Characteristics |
B10.BR strain
female
7 weeks
Cells from spleen and lymph nodes
|
| Biomaterial provider |
Jackson laboratory, Bar Harbor, Maine
|
| Treatment protocol |
Cells from spleen and lymph nodes were depleted from CD8+, MHC II+, CD11b+ and NK1.1+ cells by negative selection; stained with anti-NK1.1-PE and anti-CD4-PE-Cy5.5 and purified by FACS-sorting into a NK1.1-CD4+ T cell population (>99% purity).
|
| Growth protocol |
FACS-purified NK1.1-CD4+ T cells were activated with plate bound anti-CD3 (5ug/ml) and soluble anti-CD28 (1 ug/ml) in Bruff's medium in the presence of exogenous mIL-6 (100 ng/ml; R&D Systems) for 16 h.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated with the RNAeasy Mini Kit (Qiagen) including an on-column DNaseI-digestion according to the manufacturer's protocol.
|
| Label |
Biotin
|
| Label protocol |
Double-stranded cDNA was then synthesized from a T7-oligo(dT) primer followed by an in vitro transcription reaction (ENZO) and fragmentation reaction to produce biotin-labeled cRNA fragments that were hybridized to the probe array at 45°C for 16 hr.
|
| |
|
| Hybridization protocol |
biotin-labeled cRNA fragments that were hybridized to the genechip probe array at 45°C for 16 hr
|
| Scan protocol |
The probe arrays were scanned (Affymetrix GS3000 Scanner) to quantify the fluorescence intensity associated with each oligonucleotide probe.
|
| Description |
None
|
| Data processing |
Qspline normalization (Workman and coworkers), RMA expression statistic
(Speed and coworkers).
|
| |
|
| Submission date |
Apr 04, 2007 |
| Last update date |
Mar 18, 2009 |
| Contact name |
Mercedes Rincon |
| E-mail(s) |
mrincon@uvm.edu
|
| Phone |
1802-656-0937
|
| Fax |
1802-656-3854
|
| Organization name |
University of Vermont
|
| Department |
Medicine/Immunobiology program
|
| Street address |
89 Beaumont Ave., Given D305
|
| City |
Burlington |
| State/province |
VT |
| ZIP/Postal code |
05405 |
| Country |
USA |
| |
|
| Platform ID |
GPL8321 |
| Series (1) |
| GSE7459 |
IL-6 induced changes in gene expression in activated mouse CD4+ T cells |
|
