|
| Status |
Public on Jul 22, 2009 |
| Title |
Naïve @ 0hr, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Naive OT-I TCR transgenic CD8 T lymphocytes
|
| Organism |
Mus musculus |
| Characteristics |
cell type: CD8 T lymphocyte
genotype: OT-1 TCR transgenic
strain: C57Bl6
age: 6 to 8 weeks
naive or stimulated: naive
stimulating agent: none
time point: 0hr
biological rep: 2
|
| Treatment protocol |
At indicated times after stimulation, the cells were harvested, washed with PBS and subjected to RNA isolation procedure.
|
| Growth protocol |
CD44lo CD8+ T cells were negatively enriched from lymph nodes of OT-1 transgenic mice and stimulated in vitro with H-2Kb:SINNFEKL complex and B7-1 proteins coated on 5uM latex microspheres in presence or absence of murine rIL-12, universal type I IFN or trichostatin A. All cultures were supplemented with human rIL-2. At 0h, 24h, 48h, and 72h the cells from cultures were harvested.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Rneasy Mini Kit, Qiagen and performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10ug total RNA (Expression Analysis Technical Manual, Affymetrix) using Enzo Bioarray HighYield RNA transcript labeling kit.
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45oC on Murine GeneChip U74Av2 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400 according to Affymetrix protocol at the University of Minnesota Microarray facility.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Naïve OT-I CD8 T cells stimulated @ 0hr, biological rep2
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings as the normalization method with scaling target signal set to 1000.
|
| |
|
| Submission date |
May 01, 2009 |
| Last update date |
Nov 19, 2010 |
| Contact name |
PUJYA AGARWAL |
| E-mail(s) |
agar0050@umn.edu
|
| Phone |
612-626-2476
|
| Organization name |
University of Minnesota
|
| Department |
Laboratory,Medicine and Pathology
|
| Street address |
6-246 NHH, 312 Church Street, SE
|
| City |
Minneapolis |
| State/province |
MN |
| ZIP/Postal code |
55455 |
| Country |
USA |
| |
|
| Platform ID |
GPL81 |
| Series (1) |
| GSE15930 |
Gene expression signature of naïve and in vitro activated CD8 T cells in response to IL-12 and Type I IFN |
|
