|
Status |
Public on Jan 01, 2014 |
Title |
CD11b DC Xbp1 KO (DC conditional KO), biological replicate 1 |
Sample type |
RNA |
|
|
Source name |
CD11b, KO
|
Organism |
Mus musculus |
Characteristics |
strain: C57BL/6 tissue: Spleen genotype: Xbp1 KO
|
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 Xbp1 KO
|
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
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|
|
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 |
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