|
| Status |
Public on Jan 01, 2014 |
| Title |
CD11b DC wild type, biological replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
CD11b, WT
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
tissue: Spleen
genotype: WT
|
| Treatment protocol |
Not applicable
|
| Growth protocol |
Spleens were isolated from WT and KO mice (age around 8 weeks). Spleen was minced and digested with Liberase TM (0.02mg/ml; Roche) and Dnase I recombinant (0.01 U/µl; Roche) for 30 min at 37°C in HBSS followed by lysis of red blood cells. CD3+ and CD19+ cells were depleted using anti-CD3 FITC and anti-CD19 FITC antibodies and anti-FITC microbeads (Miltenyi Biotec). T- and B-depleted splenocytes were stained with CD11c-PE Texas red, CD8a-PeCy5, CD11b-PeCy7, MHCII-APC-Cy7, CD24-Pacific blue and live/dead marker conjugated to Amcyan. CD8+ cDCs were isolated by cell sorting gating on FSC/SSC/singlets/alive/FITC-/ MHCII+/ CD24hi/ CD8+/ CD11cint-hi whereas CD11b+ cDCs were isolated by gating on FSC/ SSC/ singlets/ alive/FITC-/ MHCII+/ CD24lo-int/ CD11b+/ CD11chi. Cell sorting was performed on a FACSAria (BD Biosciences).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNEasy Plus Micro Kit (QIAGEN)
|
| Label |
Biotin
|
| Label protocol |
Affymetrix WT Terminal Labeling Kit
|
| |
|
| Hybridization protocol |
Standard Affymetrix protocol. cDNA was hybrised on Affymetrix GeneChip Mouse Gene 1.0 ST Arrays (GPL6246).
|
| Scan protocol |
Affymetrix Gene ChIP Scanner 3000 7G
|
| Description |
Splenic CD11b DC from WT
|
| Data processing |
Data were processed using Bioconductor. Normalisation was done by RMA.
probe_group_file: MoGene-1_0-st-v1.r4.pgf
meta-probeset_file: MoGene-1_0-st-v1.r4.mps
|
| |
|
| Submission date |
Dec 02, 2013 |
| Last update date |
Jan 01, 2014 |
| Contact name |
Sophie Janssens |
| E-mail(s) |
sophie.janssens@irc.vib-ugent.be
|
| Organization name |
University of Ghent-VIB
|
| Department |
VIB Inflammation Research Center
|
| Street address |
Technologiepark 927
|
| City |
Ghent |
| ZIP/Postal code |
9000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE52857 |
Expression data in splenic DC subsets in wild type and Xbp1 deficient mice |
|
