|
| Status |
Public on Dec 04, 2012 |
| Title |
NOD.G9C8 tg, replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
mouse spleen
|
| Organism |
Mus musculus |
| Characteristics |
cell type: activated CD8 T cells
sample type: transgenic CD8 T cells from NOD mice
host genetic background: NOD/LtJ
|
| Treatment protocol |
Samples were peptide stimulated for 3 days and rested in IL2 (5 IU/ml) for 2 days. Cells were then harvested, washed with PBS and RNA was extracted.
|
| Growth protocol |
3 separate vials of frozen G9C8 clone T cells or 2 separate (4-8 wk old) transgenic mouse splenocytes were used for in vitro antigenic stimulation with 10ug/ml Insulin B peptide(LYLVCGERG).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared from 8 ug total RNA according to the standard Affymetrix (Affymetrix, Santa Clara, CA) protocol and protocols provided by the Functional Genomics Core facility (University of Chicago).
|
| |
|
| Hybridization protocol |
12ug of labeled and fragmented cRNA were hybridized to Affymetrix Mouse 430v2.0 GeneChips.GeneChips were washed and stained in the Affymetrix Fluidics station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000
|
| Description |
Gene expression from activated and rested transgenic CD8 T cells
|
| Data processing |
Affymetrix GCOS software was used to run the Comparison analysis method using default parameters, with signal scaling (target value=150) to normalize signal from each chip. Resulting files were subjected to further analysis using GeneSpring GX7.3 (Agilent Technologies) and Ingenuity pathway analysis softwares.
|
| |
|
| Submission date |
Aug 29, 2011 |
| Last update date |
Dec 04, 2012 |
| Contact name |
Alexander Chervonsky |
| Organization name |
University of Chicago
|
| Department |
Pathology
|
| Street address |
924 E 57th Street
|
| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60637 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE31726 |
Expression analysis of G9C8 clone and NOD.G9C8 tg CTLs |
|
