|
| Status |
Public on Nov 01, 2015 |
| Title |
A.E7-stim for 6hr-U74B2v |
| Sample type |
RNA |
| |
|
| Source name |
A.E7 T cells stimulated for 6hrs
|
| Organism |
Mus musculus |
| Characteristics |
tissue: T cell clone
genotype: murine B10.A origin
treatment: anti-TCR antibody
time point: 6hrs
|
| Treatment protocol |
5x10^6 cells were added to each well of the 6-well plate pre-coated with anti-TCR antbody. After 0, 2, 4 and 6hrs at 37C, 5% CO2, cells were harvested by scraping and frozen at -70C until RNA extraction. For the maintenance phase sample, cells were stimulated for 16hrs, washed three times, and incubated in fresh complete media at 37C, 5% CO2 without further stimulation for 5days before cell harvest and RNA extraction.
|
| Growth protocol |
Cells were stimulated with whole pigeon cytochrome c protein in the presence of irradiated (3000 R) B10.A splenocytes as a source of APC. After 48 h, the cells were expanded in IL-2 (10U/ml) containing complete medium and rested for at least 2 weeks before restimulation or experimental use.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using RNeasy kit(Qiagen) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 9.5 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip U74 A then on B and last on C Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
cRNAs from AE.7 cells stimulated for 6hrs were hybridized on mouse genome U74B(version2) chip.
|
| Data processing |
The data were analyzed with Microarray Suite version 4.0 (MAS 4.0) using Affymetrix default analysis settings and global scaling as normalization method. The target intensity of each array was arbitrarily set to 150.
|
| |
|
| Submission date |
Sep 04, 2015 |
| Last update date |
Nov 01, 2015 |
| Contact name |
Kyungho Choi |
| Organization name |
Seoul National University College of Medicine
|
| Department |
Department of Biochemistry and Molecular Biology
|
| Lab |
Lab of Molecular Immunology
|
| Street address |
103 Daehak-ro, Jongno-gu
|
| City |
Seoul |
| ZIP/Postal code |
110-799 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL82 |
| Series (1) |
| GSE72731 |
Kinetic analysis of T cell anergy induction |
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