Comment[ArrayExpressAccession]	E-MTAB-1816
MAGE-TAB Version	1.1
Investigation Title	MAZR and mast cells
Comment[Submitted Name]	MAZR and mast cells
Experiment Description	Mast cells are known to be the key players in type I hypersensitivity reactions in humans and mice. They are critically involved in the development of allergic rhinitis, allergic asthma and systemic anaphylaxis. In this study we investigated the role of the transcriptional regulator MAZR in mast cells by comparing the expression profile of mast cells generated from wild-type (MazrF/F) and MAZR-deficient (MazrF/F x Vav-iCre) bone marrow cells. Our results from the array data demonstrate that MAZR acts preferentially as a transcriptional repressor in mast cells.
Experimental Design	development or differentiation design	in_vitro_design	co-expression_design	genotype design
Comment[AEExperimentType]	transcription profiling by array
Comment[AEExperimentDisplayName]	Transcription profiling by array of mast cells generated from Mus musculus wild-type (MazrF/F) and MAZR-deficient (MazrF/F x Vav-iCre) bone marrow cells
Experimental Factor Name	genotype	treatment
Experimental Factor Type	genotype	treatment
Quality Control Type	biological replicate
Quality Control Term Source REF	EFO
Public Release Date	2013-10-18
Person Last Name	Ellmeier
Person First Name	Wilfried
Person Mid Initials
Person Email	wilfried.ellmeier@meduniwien.ac.at
Person Phone
Person Address	Latarettgasse 19, 1090 Vienna, Austria
Person Affiliation	Medical University of Vienna, Center for Pathophysiology, Infectiology and Immunology, Institute of Immunology
Person Roles	submitter
PubMed ID
Publication Author List	Anastasia Abramova, Shinya Sakaguchi, Alexandra Schebesta, Hammad Hassan, Nicole Boucheron, Peter Valent, Axel Roers, Wilfried Ellmeier
Publication Title	The Transcription Factor MAZR Preferentially Acts as a Transcriptional Repressor in Mast Cells and Plays a Minor Role in the Regulation of Effector Functions in Response to FcεRI Stimulation
Publication Status	in preparation
Protocol Name	P-MTAB-34136	P-MTAB-34137	P-MTAB-34138	P-MTAB-34139	P-MTAB-34140	P-MTAB-34141
Protocol Type	growth protocol	treatment protocol	nucleic acid extraction protocol	nucleic acid labeling protocol	nucleic acid hybridization to array protocol	array scanning protocol
Protocol Description	Bone marrow cells were cultured at a concentration of 0.5x106/ml in medium [RPMI (Sigma), 10% heat-inactivated FBS (PAA), 5uM B-mercaptoethanol (Gibco), penicillin/streptomycin (Gibco), 2 mM Glutamin (Gibco)] supplemented with 5 ng/ml of recombinant murine IL-3 (Peprotech, UK). Once per week, in the course of 4-5 weeks, cells were spun down, counted and re-seeded at the density of 0.5x106/ml. The purity of the BMMC population was monitored by FACS and cells were only used for assays if the c-kit+Fc_RI+ mature BMMC constituted >95% of the total cultured cells.	Priming and activation of mast cells: Mature BMMC (>95% c-kit+Fc_RI+) were primed (sensitized) in the following way: cells were spun down, counted and re-suspended at the concentration of 1x106/ml in full mast cell medium containing 1ug/ml of anti-TNP IgE mAb (#557079, BD Pharmingen) overnight. 6 well plates were coated overnight at 4C with 5 ml of 500 ng/ml of TNP-BSA in PBS, or just PBS for non-activated samples. TNP-BSA-coated plates were washed 3x in PBS, then blocked for 2 hours in 3% detoxified BSA at RT followed by 3 more washes in PBS. IgE-primed BMMCs (20x106 in 5 ml medium) were incubated on PBS or TNP-BSA coated plates for 4 hours at 37C incubator. For non-activated conditions we generated 3 (WT) and 4 (KO) independent biological replicas. For activated conditions we generated 4 (WT) and 3 (KO) independent biological replicas. 	RNA was isolated from IgE-primed or activated mast cells using 1 ml Triazol. 	Agilents Low Input Quick Amp Labeling Kit (Cy3)	The amplified cyanine 3-labeled cRNA samples were then purified using Promegas SV Total RNA Isolation System and hybridized to Agilent Whole Mouse Genome Microarrays, 8x60K according to the standard protocol of the manufacturer.	Microarray slides were washed and scanned with an Agilent Scanner, according to the standard protocol of the manufacturer.
Protocol Term Source REF	EFO	EFO	EFO	EFO	EFO	EFO
Term Source Name	EFO
Term Source File	http://www.ebi.ac.uk/efo
SDRF File	E-MTAB-1816.sdrf.txt
Publication DOI	10.1371/journal.pone.0077677