|
Status |
Public on Nov 04, 2013 |
Title |
WT Dermis CD11b+ DC, steady-state, replicate 3 |
Sample type |
RNA |
|
|
Source name |
BMN_BH4_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.
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|
|
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 |