|
| Status |
Public on Jun 25, 2013 |
| Title |
KO_Tet_1 |
| Sample type |
RNA |
| |
|
| Source name |
D54 PI
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Spleen
genotype: FoxO1-/-
cell population: CD8 CD44 high Tetramer positive 1
cell type: purified CD8
age: 6-8 weeks-old
|
| Treatment protocol |
6 to 8 week old WT and FoxO1-/- mice were infected intraperitoneally with 2 x 10^5 PFU Armstrong strain of LCMV to induce acute infection. At day 54 post infection spleens were harvested and CD8 T cells were purfied.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the sorted cells by TRIzol Reagent (per manufacturer's protocol recommendations). SuperAmp RNA amplification was performed according to Miltenyi Biotec's undisclosed procedure. Briefly, though, he amplification is based on a global PCR protocol using mRNA-derived cDNA. MRNA was isolated via magnetic bead technology and amplified cDNA samples were quantified using the ND-1000 Spetrophotometer.
|
| Label |
Cy3
|
| Label protocol |
250ng of each of the cDNAs were used as template for Cy3 labeling which was perfomed according to Miltenyi Biotec's undisclosed protocol.
|
| |
|
| Hybridization protocol |
Cy3 labeled cDNAs were hybridized overnight (17 hours, 65°C) to an Agielent Whole Mouse Genome Oligo Microarray (8 x 60K) using Agilent's recommended bybridization chamber and oven. Microarrays were washed once with Agilent Gene Expression Wash Buffer 1 for 1 min at room temperature. Followed by a second wash with preheated Agilent Gene Expression Was Buffer 2 (37°C) for 1 min. The last washing step was perfomed with acetonitrile.
|
| Scan protocol |
Fluorescence signals of the hybridized Agilent Microarrays were detected using Agilent's Microarray Scanner System (Agilent Techonologies) using one color scan settings for 8x60k array slides.
|
| Description |
Gene expression in purified CD8 D54 PI LCMV-Arm
|
| Data processing |
Agilent Feature Extraction Software was used to read out and process the microarray image files.
|
| |
|
| Submission date |
Apr 01, 2013 |
| Last update date |
Jun 25, 2013 |
| Contact name |
Melba Marie Tejera |
| Organization name |
UW-Madison
|
| Department |
Pathobiological Sciences
|
| Lab |
M. Suresh
|
| Street address |
2015 Linden Drive
|
| City |
Madison |
| State/province |
WI |
| ZIP/Postal code |
53562 |
| Country |
USA |
| |
|
| Platform ID |
GPL13912 |
| Series (1) |
| GSE45673 |
FoxO1 Controls Effector to Memory Transition and Maintenance of Functional CD8 T-Cell Memory |
|
