Comment[ArrayExpressAccession]	E-GEOD-57061
MAGE-TAB Version	1.1						
Public Release Date	2014-07-30
Investigation Title	Expression data for Lck-Cre, Med23flox/flox and Med23flox/flox;Lck-Cre thymocytes +/- 3hr exposure to plate bound anti-CD3 antibody						
Comment[Submitted Name]	Expression data for Lck-Cre, Med23flox/flox and Med23flox/flox;Lck-Cre thymocytes +/- 3hr exposure to plate bound anti-CD3 antibody
Experiment Description	MED23, a subunit of the Mediator coactivator complex, is important for the expression of a subset of MAPK/ERK pathway-dependent target genes; however, the genes in this subset varies between cell types.  MAPK/ERK pathway-dependent processes are essential for T-cell development and function, but whether MED23 has a role in this context is unknown.  We generated Med23 conditional knockout mice and induced Med23 deletion in early T cell development using the lineage specific Lck-Cre transgene.  While the total cell number and distribution of cell populations in the thymuses of Med23flox/flox;Lck-Cre mice were essentially normal, MED23 null T-cells failed to efficiently populate the peripheral lymphoid organs.  MED23 null thymocytes displayed decreased expression of the MAPK/ERK-responsive genes Egr1, Egr2, as well as of the membrane glycoprotein Cd52 (CAMPATH-1).  MED23 null CD4 single-positive thymocytes also showed decreased expression of KLF2 (LKLF), a T cell master regulatory transcription factor.  Indeed, similarities between the phenotypes of mice lacking MED23 or KLF2 in T-cells suggest that KLF2 deficiency in MED23 null T-cells is one of their key defects.  Mechanistic experiments using MED23 null MEFs further suggest that MED23 is required for full activity of the MAPK-responsive transcription factor MEF2, which has previously been shown to mediate Klf2 expression.  In summary, our data indicate that MED23 has critical roles in enabling T-cells to populate the peripheral lymphoid organs, possibly by potentiating MEF2-dependent expression of the T-cell transcription factor KLF2. 12 samples, 2 of each genotype (Lck-Cre, Med23flox/flox and Med23flox/flox;Lck-Cre) both with mock and anti-CD3 treatment						
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl					
Person Last Name	Brindle	Brindle	Kasper				
Person First Name	Paul	Paul	Lawryn				
Person Mid Initials		K	H				
Person Email	paul.brindle@stjude.org						
Person Affiliation	St Jude Children's Research Hospital						
Person Address	Biochemistry, St Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN, USA						
Person Roles	submitter						
Protocol Name	P-GSE57061-1	P-GSE57061-5	P-GSE57061-6	P-GSE57061-2	P-GSE57061-3	P-GSE57061-4	P-GSE57061-7
Protocol Description	The data were analyzed with GeneChip Operating Software version 1.4 using the default MAS 5.0 algorithm settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 500. ID_REF =  VALUE = MAS5.0 signal intensity	Biotinylated cRNA were prepared from 10 micrograms of total RNA according to the standard Affymetrix protocol (Expression Analysis Technical Manual 701025 rev1, 2001, Affymetrix)	Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on an Affymetrix Murine Genome U74A Version 2 array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.	Thymuses were harvested from mice sacrificed by CO2 inhalation and placed in DPBS+2%FBS on ice. Single cell suspensions were made by pressing the thymus through a 70um mesh strainer using the rubber plunger end from a 3cc plastic syringe and 6ml warm RPMI+10%FBS/P/S/l-glu/BME. Cells were counted and seeded at 3x10^7 per single well of a 6 well plate in 3ml final volume of same medium and allowed to rest in 37C incubator for four hours. After rest period, the thymocytes and medium were transferred to a new well +/-plate bound anti-CD3 for three hours.	Mice were maintained under protocols in accordance with St Jude IACUC	Following treatment, the cells were spun down and resuspended in 1ml of Trizol. RNA was extracted following the manufacturers instructions.	GeneChips were scanned using the AffymetrixGeneChip Scanner 3000.
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	sample treatment protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol
Experimental Factor Name	GENOTYPE						
Experimental Factor Type	genotype						
Publication Title	T-Cells Null for the MED23 Subunit of Mediator Express Decreased Levels of KLF2 and Inefficiently Populate the Peripheral Lymphoid Organs.						
Publication Author List	Kasper LH, Fukuyama T, Brindle PK						
PubMed ID	25054639						
Publication DOI	10.1371/journal.pone.0102076						
Comment[SecondaryAccession]	GSE57061						
Comment[GEOReleaseDate]	2014-07-30						
Comment[ArrayExpressSubmissionDate]	2014-04-24						
Comment[GEOLastUpdateDate]	2014-07-30						
Comment[AEExperimentType]	transcription profiling by array						
SDRF File	E-GEOD-57061.sdrf.txt