|
| Status |
Public on May 03, 2012 |
| Title |
FcR II KO unstimulated replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
FcR II KO 2 medium
|
| Organism |
Mus musculus |
| Characteristics |
treatment: unstimulated
cell type: bone-marrow derived dendritic cells
genetic background: C57Bl/6
genotype: Fcγ receptor IIb KO
|
| Treatment protocol |
Bone marrow-derived dendritic cells (BM-DCs) were transferred to new culture dishes at a density of 1 million BM-DC per 2 ml medium. The BM-DCs were incubated with either medium (unstimulated) or preformed Ag-Ab complexes (IC) containing 1 µg/ml OVA (Worthington) and 50 µg/ml rabbit anti-OVA IgG (ICN biomedicals) at 37 degrees for 4 hours.
|
| Growth protocol |
Bone marrow was harvested from the femurs and tibia derived from two female and two male mice of each mouse strain. The cells of all 12 mice were cultured separately in IMDM supplemented with GM-CSF containing supernatant in petri dishes. Non-adherent and semi-adherent cells were transferred to new dishes and fresh medium on days 4 and 7. On day 10, the cells were harvested and the % of immature CD11c+ DCs was determined by flow cytometry, which was typically above 85%.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cells by Qiagen RNA Easy kit. RNA was amplified by using the Illumina RNA Amplification kit.
|
| Label |
Biotin
|
| Label protocol |
Illumina RNA amplfication kit includes incorporation of biotin.
|
| |
|
| Hybridization protocol |
Samples were hybridised on Illumina mouse WG-6 V1.1 expression BeadChips.
|
| Scan protocol |
BeadChips were scanned on the Illumina BeadArray 500GX Scanner.
|
| Description |
04_D
|
| Data processing |
Raw data was exported by Illumina BeadStudio V3.0 software and imported and stored in Rosetta Resolver software.
Not Normalized data sheet contains intensities corrected for background by Rosetta's default error model; Normalized data sheet contains intensities normalized with Rosetta's default error model for Illumina arrays on a back-transformed linear scale and where different probes for the same gene are averaged and thus get the same value
|
| |
|
| Submission date |
Nov 15, 2011 |
| Last update date |
May 03, 2012 |
| Contact name |
Peter A.C. 't Hoen |
| E-mail(s) |
p.a.c.hoen@lumc.nl
|
| Phone |
+31 71 5269421
|
| Fax |
+31 71 5268285
|
| URL |
http://www.humgen.nl
|
| Organization name |
Leiden University Medical Center
|
| Department |
Center for Human and Clinical Genetics
|
| Street address |
PO Box 9600
|
| City |
Leiden |
| ZIP/Postal code |
2300 RC |
| Country |
Netherlands |
| |
|
| Platform ID |
GPL6481 |
| Series (1) |
| GSE33718 |
Genome-wide analysis of Fcγ receptor ligation on dendritic cells from wild-type, FcγRIIb KO or FcR γ chain KO mice |
|
