|
| Status |
Public on Oct 07, 2014 |
| Title |
CD4_TOL_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
MACS sorted CD4+ T cells from orally tolerised mice
|
| Organism |
Mus musculus |
| Characteristics |
tissue: mesenteric lymph nodes
genotype: DO11.10 RAG2-/- BALB/c
age: 8-10 weeks
cell type: CD4+ T
|
| Treatment protocol |
The study included 5 experimenta groups: Unimmunised mice, mice primed with CFA/OVA (100ug/mouse), mice primed and treated with control IgG (10mg/kg), mice primed and treated with abatacept (10mg/kg), and orally tolerised mice that were generated by feeding with OVA in the drinking water (50mg/ml). For CD4+ T cells draining LNs were used (unimmunised: pooled LNs and spleeen, primed: axillary, orally tolerised: mesenteric LNs) CD4+ T cells were isolated using MACS negative selection with over 90% purity. CD11c+ cells were isolated from pooled LNs and spleen using CD11c microbeads and MACS with over 90% purity.
|
| Growth protocol |
DO11.10 RAG2-/- mice were kept in ivc cages under home office regulation
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total mRNA was isolated using the RNEASY mini kit (Qiagen, UK) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
50 ng of total RNA was amplified and labeled with Ovation® Pico WTA System and Encore™ Biotin Module assay (NuGEN Technologies Inc , San Carlos, CA), target labeling process was performed according to the manufacturer's recommendation.
|
| |
|
| Hybridization protocol |
2.5 ug of labeled cDNA was hybridized on Affymetrix GeneChip HT MG-430 PM array according to the manufacturer’s recommendations.
|
| Scan protocol |
GeneChips were scanned using Affymetrix HT Array Plate and scanned images were subjected to visual inspection and a chip quality report was generated by Expression console (Affymetrix).
|
| Description |
Gene expression data from CD4+ T cells from orally tolerised mice
|
| Data processing |
The data were analyzed with the robust multi-array analysis (RMA) algorithm, obtained from www.bioconductor.org
|
| |
|
| Submission date |
Oct 06, 2014 |
| Last update date |
Oct 07, 2014 |
| Contact name |
Agapitos Patakas |
| E-mail(s) |
[email protected]
|
| Phone |
+441413305015
|
| Organization name |
University of Glasgow
|
| Department |
Institute of Infection, Immunity and Inflammation
|
| Lab |
GBRC
|
| Street address |
120 University Place
|
| City |
Glasgow |
| State/province |
Lanarkshire |
| ZIP/Postal code |
G12 8TA |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL11180 |
| Series (1) |
| GSE62089 |
Expression data from CD4+ and CD11c+ cells from DO11.10 RAG2-/- mice |
|
