Comment[ArrayExpressAccession]	E-GEOD-48934
MAGE-TAB Version	1.1					
Public Release Date	2013-07-16
Investigation Title	MicroRNA profiling of brain metastasis competent cell-derived exosomes					
Comment[Submitted Name]	MicroRNA profiling of brain metastasis competent cell-derived exosomes
Experiment Description	Objective of this work was to characterize the miRNA profile of exosomes isolated from brain-homing cell lines (MDA-MB-231BR, CTC1BMSM, and 70W) with their respective parental non-brain metastatic cell lines (MDA-MB-231P, CTC1P and MeWo). Exosomes derived from the six cell lines were isolated. Brain metastatic cell-derived exosomal miRNAs were compared with non brain metastatic cell lines (MDA-MB-231BR versus MDA-MB-231P, CTC1BMSM versus CTC1P, and 70W versus MeWo).					
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	Camacho	Camacho	Marchetti			
Person First Name	Laura	Laura	Dario			
Person Email	laura_mond@hotmail.com					
Person Affiliation	Baylor College of Medicine					
Person Address	Baylor College of Medicine, One Baylor Plaza, Houston, USA					
Person Roles	submitter					
Protocol Name	P-GSE48934-1	P-GSE48934-4	P-GSE48934-5	P-GSE48934-2	P-GSE48934-3	P-GSE48934-6
Protocol Description	The normalization and all the data analysis were performed according to the manufacturers instructions using their web-based software package: http://pcrdataanalysis.sabiosciences.com/mirna/arrayanalysis.php. The relative quantity of the target miRNA was normalized using Syn-cel-miR-39 miScript miRNA Mimic miRNeasy Spike-In Control (Qiagen, Germantown, MD) that was exogenously added during RNA purification. The web-based software package automatically performs all deltadeltaCt based fold-change calculations from the uploaded raw threshold cycle data. Fold-Change file reports test/control (ie, brain metastatic cell-derived exosomes/non brain metastatic cell derived-exosomes).  Linked as supplementary file on Series record. ID_REF =  VALUE = normalized signal (against cel-miR-39)	10 ng of total RNA were reverse transcribed with miScript II RT kit (Qiagen, Germantown, MD) according to manufacturersM-bM-^@M-^Y guidelines. 20 M-5l of cDNA was diluted with RNase-free water prior to the PCR reaction. Real-time PCR for mature miRNA expression profiling was developed using the SYBR Green-based human breast cancer pathway-focused miScript miRNA PCR array (Qiagen, Germantown, MD) on a StepOnePlus 96-well RT PCR instrument (Applied Biosystems, Grand Island, NY).	n/a	After 72 hours of cell culturing, 10 ml of culture media was centrifuged at 3000 x g for 15 min to remove cells and cell debris. The supernatant was mixed with 2 ml of ExoQuick-TC, refrigerated for 16 hours and then centrifuged at 1500 x g for 30 min at 4M-0C to obtain the exosomes pellet.	Total RNA from exosomes was isolated using mirVana miRNA isolation kit (Life Technologies, Grand Island, NY)	n/a
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol
Experimental Factor Name	BRAIN-HOMING CELL LINE	PARENTAL NON-BRAIN METASTATIC CELL LINE	CELL TYPE			
Experimental Factor Type	brain-homing cell line	parental non-brain metastatic cell line	cell type			
Publication Title	MicroRNA and protein profiling of brain metastasis competent cell-derived exosomes.					
Publication Author List	Camacho L, Guerrero P, Marchetti D					
PubMed ID	24066071					
Publication DOI	10.1371/journal.pone.0073790					
Comment[SecondaryAccession]	GSE48934					
Comment[GEOReleaseDate]	2013-07-16					
Comment[ArrayExpressSubmissionDate]	2013-07-16					
Comment[GEOLastUpdateDate]	2013-10-15					
Comment[AEExperimentType]	transcription profiling by RT-PCR					
Comment[AdditionalFile:Data1]	GSE48934_fold-change.txt					
Comment[AdditionalFile:Data2]	GSE48934_non-normalized_data.txt					
SDRF File	E-GEOD-48934.sdrf.txt