|
| Status |
Public on May 29, 2019 |
| Title |
Memory t3 Treated Stimulated Donor 2 |
| Sample type |
RNA |
| |
|
| Source name |
Memory CD4+ T-cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Memory CD4+ T-cells
donor: 2
il-6/sil-6r: Treated
anti-cd3/anti-cd28: Stimulated
time point: t3
|
| Treatment protocol |
CD4+ T-cells were treated with indicated final concentrations of IL-6 and equimolar sIL-6R (both PeproTech EC) in a 1ml total volume (24 well plate) or 0.5ml volume (48 well plate) for 72 hours at 37°C with 5% CO2. Cells were harvested and washed twice, to remove IL-6/sIL-6R, before stimulation for 3-6 days with plate bound anti-CD3 (0.2μg/ml or 0.5μg/ml; eBioscience) and 1μg/ml soluble anti-CD28 (Biolegend).
|
| Growth protocol |
Naïve and memory CD4+ T-cells were isolated using a RosetteSep Human CD4+ T-cell Enrichment Cocktail (Stemcell Technologies) followed by CD45RO MicroBeads (MiltenyiBiotech) and cultured in serum-free medium (TexMACS, Miltenyi Biotech).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted from naïve and memory CD4+ T-cells at indicated experimental timepoints using RNeasy Mini kit (Qiagen), as per manufacturer’s instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylation of cRNA
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
200627770001_G
200627770001_G_Grn.idat
72h IL6 treated and 144h TCR stimulated cells
|
| Data processing |
Background correction, transformation and normalization were performed using bgAdjust, VST and RSN Lumi R package methods, respectively.
|
| |
|
| Submission date |
May 28, 2019 |
| Last update date |
Oct 10, 2019 |
| Contact name |
Arthur Pratt |
| E-mail(s) |
arthur.pratt@newcastle.ac.uk
|
| Organization name |
Newcastle University
|
| Department |
Faculty of Medical Sciences
|
| Lab |
Musculoskeletal Research Group
|
| Street address |
Framlington Place
|
| City |
Newcastle upon Tyne |
| ZIP/Postal code |
NE2 4HH |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL10558 |
| Series (1) |
| GSE131866 |
IL-6 mediated transcriptional programming of CD4+ T cells in early rheumatoid arthritis |
|
