Comment[ArrayExpressAccession]	E-GEOD-45147
MAGE-TAB Version	1.1						
Public Release Date	2014-02-03
Investigation Title	Hemangioblast skewing from embryonic stem(ES) cells by defined factors						
Comment[Submitted Name]	Hemangioblast skewing from embryonic stem(ES) cells by defined factors
Experiment Description	Successful derivation of a specific cell lineage from pluripotent stem cells will tremendously facilitate the clinical usage of pluripotent stem derived somatic cells. Herein, we demonstrate that ER71/Etv2, GATA2 and Scl form a core network in hemangioblast development and that transient co-expression of these three factors robustly induced hemangioblasts from ES cells. Such induced hemangioblasts potently generated hematopoietic and endothelial cells in culture as well as in vivo, warranting the evaluation of these cells in the future for repairing and/or regenerating hematopoietic and/or angiogenic defects. We have established a doxycycline inducible ES cell, iEGS, in which ER71/Etv2, GATA2 three transcription factors can be transiently co-expressed by doxycycline induction. We further analyzed the downstream target genes and signaling pathways at 6, 12 and 24hrs after ER71/Etv2, GATA2 induction. These data were obtained from three independent experiments.						
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	Liu	Yu	Liu	Choi			
Person First Name	Fang	Jinsheng	Fang	Kyunghee			
Person Email	fliu@pathology.wustl.edu						
Person Affiliation	Washington University in St. Louis						
Person Phone	314-362-9045						
Person Address	Pathology & Immunology, Washington University in St. Louis, 660S Euclid Ave., St. Louis, MO, USA						
Person Roles	submitter						
Protocol Name	P-GSE45147-1	P-GSE45147-5	P-GSE45147-6	P-GSE45147-2	P-GSE45147-3	P-GSE45147-4	P-GSE45147-7
Protocol Description	Data were processed using Affymetrix Expression Console software. Gene level expression were summarized from the intensities and then normalized using the Robust Multiarray Analysis (RMA) algorithm. probe_group_file:  MoGene-1_1-st-v1.r4.pgf meta-probeset file:  MoGene-1_1-st-v1.r4.mps ID_REF =  VALUE = RMA normalized linear gene level expression values from Affymetrix Expression Console	Biotinylated cDNA were prepared according to the standard NuGen WT-Ovation Pico RNA Amplification system from 500pg-50 ng total RNA.	Following fragmentation with NuGen Encore Biotin Module, 2.5 ug of cDNA were hybridized on Exon 1.0 ST Array in the GeneChip Hybridization Oven 640 for 18 hr at 45C. GeneChip were washed and stained in the Affymetrix Fluidics Station 450.	Doxycycline (1M-5g/ml) was added in iEGS+DOX group from day 2 to 3.	Mouse ES cells were maintained on PMEF cells in DMEM with 15% pre-selected fetal calf serum (FCS), 2% LIF, 1% L-glutamine, 1% NEAA and 4.5x 10M-bM-^@M-^S4 M MTG. ES cells were differentiated in IMDM containing 15% differentiation-screened FCS, 1% L-glutamine, 50 g/ml ascorbic acidand 4.5x 10M-bM-^@M-^S4 M MTG.	Total RNA were extracted and purified by RNeasy Mini Kit (Qiagen.) cDNA synthesis and amplification was performed using the WT cDNA synthesis and amplification kit (Affymetrix).	GeneChips were scanned using the Affymetrix GeneChip Scanner 7G 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	TREATMENT	REPLICATE	TIME	GENOTYPE			
Experimental Factor Type	treatment	replicate	time	genotype			
Publication Title	Enhanced hemangioblast generation and improved vascular repair and regeneration from embryonic stem cells by defined transcription factors.						
Publication Author List	Liu F, Bhang SH, Arentson E, Sawada A, Kim CK, Kang I, Yu J, Sakurai N, Kim SH, Yoo JJ, Kim P, Pahng SH, Xia Y, Solnica-Krezel L, Choi K						
PubMed ID	24052951						
Publication DOI	10.1016/j.stemcr.2013.06.005						
Comment[SecondaryAccession]	GSE45147						
Comment[GEOReleaseDate]	2014-02-03						
Comment[ArrayExpressSubmissionDate]	2013-03-13						
Comment[GEOLastUpdateDate]	2014-02-04						
Comment[AEExperimentType]	transcription profiling by array						
SDRF File	E-GEOD-45147.sdrf.txt