|
| Status |
Public on Jan 09, 2014 |
| Title |
naïve B cells CD40L alone; AD-HIES pt 3 |
| Sample type |
RNA |
| |
|
| Source name |
naïve B cells from AD-HIES patients
|
| Organism |
Homo sapiens |
| Characteristics |
diagnosis: Autosomal dominant hyper IgE syndrome (AD-HIES)
cell type: naïve B cells
|
| Treatment protocol |
B cells were stimulated with recombinant CD40 ligand alone or together with IL-21 (50 ng/mL)
|
| Growth protocol |
Human naïve B cells were cultured in RPMI1640 containing 10% FCS for 4 days
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
The Affymetrix GeneChip Whole Transcript Sense Target Labelling Assay was performed to generate amplified and biotinylated sense-strand DNA targets from mRNA. Briefly, cDNA was synthesised and amplified from RNA samples, then antisense RNA (cRNA) transcribed. The transcribed cRNA was then purified and used as a template for the synthesis of single-stranded DNA in the sense orientation. The sense strand DNA underwent further purification, fragmentation and labelling steps according to the Affymetrix GeneChip® Whole Transcript Sense Target Labelling Assay protocol.
|
| |
|
| Hybridization protocol |
Hybridization was performed using the Affymetrix GeneChip® Hybridization, Wash and Stain kit.
|
| Scan protocol |
Scanning was performed by the Ramaciotti Centre for Gene Function Analysis (University of New South Wales, Australia) according to the Affymetrix GeneChip® Expression Wash, Stain and Scan User Manual (P/N 702731).
|
| Description |
RNA from AD-HIES patient naïve B cells stimulated with CD40L alone
|
| Data processing |
Microarray data was analysed using GenePettern software (v3.2.3, Broad Institute). A custom contrast was then created to display genes with increased expression in the presence of IL-21 in normal B cells, but not in STAT3 deficient B cells
|
| |
|
| Submission date |
Oct 23, 2013 |
| Last update date |
Sep 01, 2016 |
| Contact name |
Stuart Tangye |
| E-mail(s) |
s.tangye@garvan.org.au
|
| Organization name |
Garvan Institute for Medical Research
|
| Department |
Immunology
|
| Street address |
384 Victoria St
|
| City |
Darlinghurst |
| State/province |
NSW |
| ZIP/Postal code |
2010 |
| Country |
Australia |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE51587 |
Identification of IL-21-induced STAT3 dependent genes in human B cells |
|
| Relations |
| Reanalyzed by |
GSE86357 |
