|
| Status |
Public on Mar 11, 2014 |
| Title |
CCR6+ CD4+ T cells_activated_2hr_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
CCR6+ CD4+ T cells_activated_2hr
|
| Organism |
Homo sapiens |
| Characteristics |
stimulation: anti-CD3 & anti-CD28 coated beads in the presence of Th17 polarizing cytokines and antibodies
|
| Treatment protocol |
Purified CCR6+ T cells were stimulated with anti-CD3 and anti-CD28 coated beads (Miltenyi) at 2:1 bead/cell ratio in the presence of Th17 polarizing cytokines and antibodies including 10ng/ml IL1b (R&D), 10ng/ml IL23 (R&D), 30ng/ml TGFb1 (R&D), g/ml anti-IL4 and anti-IFN (eBioscience)
|
| Growth protocol |
Stimulated T cells were cultured in a 24-well tissue culture plate at 5x106 cells/well in 1ml of IMDM medium containing 10% serum replacement factor, 2mM glutamine, 1mM sodium pyruvate, 10mM HEPES, 1mM MEM nonessential amino acid solution, and 100 U/ml each of penicillin G and streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with a Qiagen RNeasy kit using the manufacturer's protocol.
|
| Label |
biotin
|
| Label protocol |
Target was prepared from 50ng total RNA using NuGEN Ovation V2 kits, followed by NuGEN Encore biotin labeling module using the manufacturer's protocol.
|
| |
|
| Hybridization protocol |
All samples were hybridized in parallel to different arrays on the same HT array plate using the manufacturer's protocol. Arrays were washed and stained on an Affymetrix GeneTitan instrument.
|
| Scan protocol |
Arrays were scanned using an Affymetrix GeneTitan MC scanner.
|
| Description |
Memory CD4+ T cells purified from PBMCs through negative selection followed by positive selection with CCR6 biotin and anti-biotin magnetic beads
|
| Data processing |
Array Studio was used to perform RMA normalization from CEL files
|
| |
|
| Submission date |
Jul 17, 2013 |
| Last update date |
Mar 11, 2014 |
| Contact name |
Anton Bittner |
| E-mail(s) |
abittner@its.jnj.com
|
| Organization name |
Janssen R&D
|
| Department |
Systems Biology
|
| Street address |
3210 Merryfield Row
|
| City |
San Diego |
| State/province |
CA |
| ZIP/Postal code |
92121 |
| Country |
USA |
| |
|
| Platform ID |
GPL13158 |
| Series (1) |
| GSE48978 |
Comparison of RNA-Seq and Microarray in Transcriptome Profiling During T Cell Activation |
|
