|
| Status |
Public on Mar 06, 2014 |
| Title |
HUVEC_PPARD_normoxia_DMSO |
| Sample type |
RNA |
| |
|
| Source name |
HUVECs, DMSO, normoxia, 24h
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: human umbilical vein cells (HUVECs)
ppard treatment: DMSO
oxygen condition: normoxia
|
| Treatment protocol |
1.5x10^5 HUVEC cells were plated in 6 well culture plate, and cultivated for 2 days. They were stimulated with PPARβ/δ ligand (GW501516) at a concentration of 100 nM and/or hypoxia (1% O2) for 24 hours. Same concentration of DMSO was used as a control sample, and as to the oxgen, normoxia was used as a control condition.
|
| Growth protocol |
Human umbilical vein endothelial cells (HUVECs) (Lonza, Basel, Switzerland) were cultured in EGM2MV medium (Lonza). HUVECs were used within the first 6 passages.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were extracted using Isogen (Nippon Gene, Osaka, Japan). 3 micrograms of total RNA was reverse transcribed using the SuperScript III system for cDNA synthesis (Invitrogen) according to the protocol recommended by Affymetrix.
|
| Label |
biotin
|
| Label protocol |
In vitro transcription was performed on 1 microgram of cDNA using the Enzo BioArray High Yield RNA transcript labelling kit (Enzo Diagnostics). The cRNA was cleaned using RNeasy clean-up columns (Qiagen). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc.
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| |
|
| Hybridization protocol |
10 micrograms of fragmented cRNA were hybridised (45 degrees CelsiusC, 16 hours). Hybridization was controlled by use of the GeneChip Eukaryotic Hybridization Control Kit (Affymetrix). Washing and staining were performed in a Fluidics Station 450 (Affymetrix) using the protocol EukGE-WS2v4.
|
| Scan protocol |
Scanning was performed in an Affymetrix GeneChip scanner 3000 7G.
|
| Description |
24hrs after DMSO treatment.
|
| Data processing |
The data were analyzed with Robust Multi-array Average (RMA) using GeneSpring 12.5 default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
Aug 27, 2013 |
| Last update date |
Mar 06, 2014 |
| Contact name |
Tsuyoshi Inoue |
| E-mail(s) |
tsinoue-tky@umin.ac.jp
|
| Organization name |
The University of Tokyo
|
| Street address |
Hongo
|
| City |
Tokyo |
| ZIP/Postal code |
113-8655 |
| Country |
Japan |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE50378 |
Expression data of HUVEC cells after PPARβ/δ agonist and/or hypoxia treatment |
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