Comment[ArrayExpressAccession]	E-GEOD-44692
MAGE-TAB Version	1.1														
Public Release Date	2014-04-29
Investigation Title	MiR-155 promotes blood-brain barrier dysfunction in neuroinflammation (part 1)														
Comment[Submitted Name]	MiR-155 promotes blood-brain barrier dysfunction in neuroinflammation (part 1)
Experiment Description	Here, we investigated the time-course changes in the pattern of microRNA (miRNA) expression of TNFα and IFNγ-stimulated and unstimulated hCMEC/D3 cells,  an immortalized human cerebral microvascular endothelial cell line. In order to investigate pro-inflammatory cytokine-induced changes in miRNA  levels in hCMEC/D3 cells, we challenged brain endothelial cells with TNFα and IFNγ (100 ng/ml) for 2 h, 6 h and 24 h and determined microRNA expression in cytokine-stimulated and unstimulated cells														
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	Lopez-Ramirez	Lopez-Ramirez	Wu	Pryce	Simpson	Reijerkerk	King-Robson	Kay	de Vries	Hirst	Sharrack	Baker	Male	Michael	Romero
Person First Name	Miguel	Miguel	Dongsheng	Gareth	Julie	Arie	Josh	Oliver	Helga	Mark	Basil	David	David	Gregory	Ignacio
Person Mid Initials	Alejandro	A			E				E	C			K	J	A
Person Email	m.a.lopez-ramirez@yale.edu														
Person Affiliation	Yale University														
Person Address	Yale Cardiovascular Research Center, Yale University, 300 George street, New Haven, USA														
Person Roles	submitter														
Protocol Name	P-GSE44692-1	P-GSE44692-5	P-GSE44692-6	P-GSE44692-2	P-GSE44692-3	P-GSE44692-4	P-GSE44692-7								
Protocol Description	standard Agilent Feature Extraction protocol ID_REF =  VALUE = Normalized signal intensity	Total RNA (100 ng) was labelled with pCp-Cy3 using T4 RNA ligase (GE healthcare, Amersham, UK)	Total RNA including miRNA (100 ng) was first dephosphorylated with calf intestine alkaline phosphatase (GE healthcare, Amersham, UK), denatured with dimethyl sulfoxide, and labeled with pCp-Cy3 to the 3’-ends using T4 RNA ligase (GE healthcare, Amersham, UK). The labeled RNAs were hybridized to Agilent human miRNA microarray (Santa Clara, CA). Design ID(AMADID)=021827. Standard Agilent’s miRNA microarray hybridization protocol	hCMEC/D3 cells were seeded onto collagen-coated tissue culture plate supplied by Greiner Bio-one (Gloucestershire, UK) and maintained for 72 h after confluence. Culture media was then changed to EGM-2 media without VEGF. Cells were left untreated or treated with TNFα and IFNγ at the concentrations and times indicated for each experiment.	hCMEC/D3 cells were cultured in EGM-2 MV medium (Lonza, Slough Wokingham, UK) and supplemented with the following components obtained from the manufacturer: 0.025 % (v/v) rhEGF, 0.025 % (v/v) VEGF, 0.025 % (v/v) IGF, 0.1 % (v/v) rhFGF, 0.1 % (v/v) gentamycin, 0.1 % (v/v) ascorbic acid, 0.04 % (v/v) hydrocortisone and 2.5 % (v/v) foetal bovine serum (FBS) (hereafter referred to as EGM-2 medium).	Total RNA of hCMEC/D3 cells were isolated using miRNeasy mini kit (Qiagen, Sussex, UK) according to the manufacturer’s protocols. The quantity (NanoDrop 1000 spectrophotometer) and the quality (2100 Bioanalyzer, RNA 6000 Pico LabChip; Agilent, Palo Alto, CA, USA) of the total RNA were analyzed prior to determination of mRNA levels	Standard Agilent scaning protocol								
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	TIME													
Experimental Factor Type	treatment	time													
Comment[SecondaryAccession]	GSE44692														
Comment[GEOReleaseDate]	2014-04-29														
Comment[ArrayExpressSubmissionDate]	2013-02-26														
Comment[GEOLastUpdateDate]	2014-04-29														
Comment[AEExperimentType]	transcription profiling by array														
Comment[AdditionalFile:Data1]	GSE44692_averaged_by_mirna.txt														
Comment[AdditionalFile:Data2]	GSE44692_normalized_by_mirna.txt														
SDRF File	E-GEOD-44692.sdrf.txt