|
| Status |
Public on May 17, 2008 |
| Title |
CD8 T cells_IL-2 complex treatment_3 h |
| Sample type |
RNA |
| |
|
| Source name |
CD8 T cells_IL-2 complex treatment_3 h
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
gender: male
tissue: spleen
|
| Biomaterial provider |
Jackson Laboratory
|
| Treatment protocol |
IL-2 complex was prepared by mixing 50 ug anti-IL-2 mAb (S4B6) and 1.5 ug recombinant mouse IL-2, which was then injected i.p. into C57BL/6J mice. Total CD8 T cells from the spleen at 3 h post treatment were purified by magnetic cell sorting using the CD8a T cell isolation kit, supplemented with biotinylated anti-CD11c, anti-CD19, anti-Gr-1, anti-TCRγδ, anti-I-Ab, and anti-NK1.1 mAbs. The purity was > 95%. Total RNA from the purified CD8 T cells was subject to DNA microarray analysis using GeneChip Mouse genome 430 2.0 array. Target labeling, hybridization and scanning were performed at the Center for Array Technologies at the University of Washington.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from purified CD8 T cells using RNeasy Mini Kit (QIAGEN), then concentrated by EtOH precipitation.
|
| Label |
biotin
|
| Label protocol |
Total RNA (~4.65ug) was used to create the first strand cDNA using a T7-linked oligo(dT) primer. Once second strand synthesis was complete, an in vitro transcription reaction was performed using biotinylated UTP and CTP in the Affymetrix IVT Kit.
|
| |
|
| Hybridization protocol |
Labeled cRNA was processed further as recommended by Affymetrix, where 15 ug of cRNA was fragmented, a hybridization cocktail was assembled with addition of spike-in controls, and chips were hybridized for 16 hours. The chips were then washed and stained with streptavidin-phycoerythrin using the Affymetrix GeneChip System.
|
| Scan protocol |
The genechips were placed in the GeneChip Scanner 3000, which scans the chips at a resolution of less than 2 microns.
|
| Description |
Labeling, hybridization, and scanning were performed at the Center for Array Technologies at the University of Washington.
|
| Data processing |
Image processing and expression analysis were performed using Affymetrix GCOS software.
|
| |
|
| Submission date |
Sep 03, 2009 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Michael J Bevan |
| Organization name |
Univerisity of Washington, HHMI
|
| Department |
Immunology
|
| Lab |
Bevan
|
| Street address |
I604H HSC, 1959 NE Pacific Street
|
| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98195-7370 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE11446 |
CD8 T cells stimulated with IL-2 complex in vivo |
|
| Relations |
| Reanalyzed by |
GSE119085 |
