Comment[ArrayExpressAccession]	E-GEOD-19098
Public Release Date	2010-12-15
Investigation Title	Expression data from human umbilical vein endothelial cells (HUVECs) as a function of cell adhesion and VEGF exposure							
Comment[Submitted Name]	Expression data from human umbilical vein endothelial cells (HUVECs) as a function of cell adhesion and VEGF exposure
Experiment Description	Angiogenesis is tightly regulated by both soluble growth factors and cellular interactions with the extracellular matrix (ECM).  While cell adhesion via integrins has been shown to be required for growth factor signaling and downstream angiogenesis, the effects of quantitative changes in cell adhesion and spreading against the ECM remain less clear. Examining changes in global gene expression in limited versus high adhesion contexts in human umbilical vein endothelial cells, we demonstrated a VEGF-induced upregulation of genes associated with vascular invasion and remodeling when cell adhesion was restricted, whereas cells on highly adhesive surfaces upregulated genes associated with a proliferative response. HUVECs were cultured on micropatterned islands of fibronectin or allowed to spread fully for 2 hrs and then stimulated with 50ng/ml VEGF or no growth factor for 18 hrs in starve media.  Sample RNA from three biological replicates was extracted and prepared for hybridization on Affymetrix microarrays.							
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Term Source File	http://efo.svn.sourceforge.net/viewvc/efo/trunk/src/efoinowl/efo.owl							
Person Last Name	Shen	Shen	Raghavan	Chen				
Person First Name	Colette	Colette	Srivatsan	Christopher				
Person Mid Initials		J		S				
Person Email	geo@ncbi.nlm.nih.gov							
Person Affiliation	University of Pennsylvania							
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Person Address	University of Pennsylvania, 210 S. 33rd St., 510 Skirkanich Hall, Philadelphia, PA, USA							
Person Roles	submitter							
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Protocol Name	P-GSE19098-1	P-GSE19098-6	P-GSE19098-3	P-GSE19098-8	P-GSE19098-7	P-GSE19098-2	P-GSE19098-4	P-GSE19098-5
Protocol Description	ID_REF = <br>VALUE = GC-RMA signal (linear)	Following fragmentation, cRNA were hybridized for 16 hr at 45C on Human Affymetrix U133+ v2.0 arrays. The microarrays were then washed at low (6X SSPE) and high (100mM MES, 0.1M NaCl) stringency and stained with streptavidin-phycoerythrin.  Fluorescence was amplified by adding biotinylated anti-streptavidin and an additional aliquot of streptavidin-phycoerythrin stain.	HUVECs were cultured in full growth medium (M199+20%FBS+ 50mg/ml ECGS+100mg/ml heparin+100units/ml penicillin+100 mg/ml streptomycin) on gelatin-coated surfaces prior to treatment.	The data were analyzed using GC-RMA.	A confocal scanner was used to collect fluorescence signal at 3um resolution after excitation at 570 nm. The average signal from two sequential scans was calculated for each microarray feature.	HUVECs were cultured on micropatterned islands of fibronectin or allowed to spread fully on fibronectin for 2 hrs and then stimulated with 50ng/ml VEGF or no growth factor for 18 hrs in starve media (5% FBS in M199).	Total RNA was extrated using the Qiagen RNeasy Mini Kit according to the manufacturer's instructions.	Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 2ug total RNA.
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Protocol Type	bioassay_data_transformation	hybridization	grow	feature_extraction	image_aquisition	specified_biomaterial_action	nucleic_acid_extraction	labeling
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Comment[SecondaryAccession]	GSE19098							
Comment[GEOLastUpdateDate]	2010-12-15
Comment[AEExperimentType]	transcription profiling by array							
Comment[GEOReleaseDate]	2010-12-15
Comment[ArrayExpressSubmissionDate]	2009-11-19
SDRF File	E-GEOD-19098.sdrf.txt