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Experimental information  

Experiment series 1 : Agonists into HUVEC  

HUVEC were treated with either 50ng/ml of BMP (Bone Morphogenetic Protein)-2,
BMP-4, Fenofibrate (PPARa agonist), TGF (Tumor Growth Factor)-b, VEGF (Vascular
Endothelial cell Growth Factor), or IFN (interferon)-g for 0 hour, 1 hour and 4 hours.

Notation/SymbolOfficial Name
FenofibrateFenofibrate
BMP4Bone morphogenetic protein 4
BMP2Bone morphogenetic protein 2
TGFbTransforming growth factor, beta 1 (Camurati-Engelmann disease)
VEGFVascular endothelial growth factor
IFNgInterferon gamma
TNFaTumor necrosis factor (TNF superfamily, member 2)


Experiment series 2 : TNFa + Inhibitors into HUVEC  

HUVEC were serum-starved in EBM-2 medium containing 0.5% FBS. 18 h later, HUVEC
were pretreated for 30 min with 10uM of MG132, 50uM LY294002, 200mg/ml of
Cycloheximide, 10uM of PD98059, or 1uM of K11430 and then incubated in the
presence of 10ng/ml human TNF-alpha for 4 h. Total RNA were then extracted by
TRIzol reagent (Invitrogen, Gaithersburg, MD) according to the manufacturer's protocol.
Preparation of cRNA and hybridization of probe arrays (Human Genome U133 Plus 2.0
Array) were performed according to the manufacture (Affymetrix, Santa Clara, CA).
The results are representative of three independent experiments.

NotationSymbolOfficial NameTarget
TNFTNFTumor necrosis factor
(TNF superfamily, member 2)
-
CHXCHXCycloheximideProtein synthesis inhibitor
MGMG-132MG-132Proteasome inhibitor
PDPD98059PD98059
[2'-Amino-3'-methoxyflavone]
MAPK/ERK inhibitor
KK11430K11430VCAM-1 inhibitor
LYLY294002LY294002
[2-(4-Morpholinyl)-8-phenyl
-4H-1-benzopyran-4-one]
PI-3K inhibitor



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Copyright (c) 2005 LSBM, RCAST, The University of Tokyo All rights reserved.