|
| Status |
Public on Nov 04, 2013 |
| Title |
WT Dermis CD11b+ DC, steady-state, replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
BMN_BH2_MoGene
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
genotype/variation: WT
tissue origin: Dermis
cell type: Dermal DC
cell phenotype: CD11b+ CD24- CD64- MHCII+
inflammation: None
gender: female
age: 6 weeks-old
|
| Treatment protocol |
Inflammation was induced by application of 10µl of a 0.5% of DNFB diluted in acetone/olive oil (4:1). Ears and skin draining lymph nodes were harvested from untreated animals or 3 days post inflammation. Ear skin tissue was digested by dispase and collagenase 4 treatment, cells were enriched by percoll gradient and resuspended in PBS 2%FCS, EDTA 5mM before sorting by flow cytometry. Cutaneous lymph nodes were digested by collagenase 2 treatment, cells were enriched by optiprep gradient and resuspended in PBS 2%FCS, EDTA 5mM before sorting by flow cytometry.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from between 5x10E3 and 1x10E4 cells for each leukocyte subset with the Qiagen micro RNAeasy kit, according to the manufacturer protocol.
|
| Label |
biotin
|
| Label protocol |
Biotinylated double strand cDNA targets were prepared, from 5 ng to 20 ng of total RNA depending on the amount available per individual samples, using the NuGEN Ovation Pico WTA System V2 kit (Cat # 3302) and the NuGEN Encore Biotin Module kit (Cat # 4200) according to NuGEN recommendations.
|
| |
|
| Hybridization protocol |
Following fragmentation, 2 µg of cRNA were hybridized for 16 hr at 45°C on GeneChip Mouse Gene 1.0 ST . GeneChips were washed and stained in the Affymetrix Fluidics Station 450 .
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G.
|
| Description |
WT CD11b+ Dermal DC from steady state B6 mice, FACS sorted as: live, CD11b+ CD24- Ly6C- CD64- MHCII+
|
| Data processing |
The data were RMA normalized in R and Bioconductor, using the affy package.
|
| |
|
| Submission date |
Jul 30, 2013 |
| Last update date |
Oct 23, 2015 |
| Contact name |
Marc DALOD |
| E-mail(s) |
dalod@ciml.univ-mrs.fr
|
| Organization name |
Center of Immunology of Marseille Luminy (CIML)
|
| Department |
Dendritic cells and antiviral defense
|
| Lab |
dalod
|
| Street address |
Parc scientifique de Luminy, Case 906
|
| City |
Marseille |
| ZIP/Postal code |
13288, Cedex 09 |
| Country |
France |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE49358 |
Genome-wide expression study of the CD11b+ subsets of dermal myeloid cells and their migratory counterparts in the draining lymph node |
|
| Relations |
| Reanalyzed by |
GSE74316 |
