|
Status |
Public on May 29, 2019 |
Title |
Naive t2 Treated Unstimulated Donor 1 |
Sample type |
RNA |
|
|
Source name |
Naïve CD4+ T-cells
|
Organism |
Homo sapiens |
Characteristics |
cell type: Naïve CD4+ T-cells donor: 1 il-6/sil-6r: Treated anti-cd3/anti-cd28: Unstimulated time point: t2
|
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 |
200627770010_I 200627770010_I_Grn.idat 72h IL6 treated and 4h 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 |
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