|
| Status |
Public on Jan 01, 2013 |
| Title |
Endothelial Cells siRNA KLF2 rep2 |
| Sample type |
RNA |
| |
|
| Source name |
HUVECs_siRNA-KLF2
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: KLF2 siRNA with 4h statin
cell type: Human umbilical vein cells
|
| Treatment protocol |
1.5x105 HUVEC cells were plated in 6 well culture plate. Next day before statin treatment siRNA KLF2 or KLF4 were transfected for 1day, and then HUVECs were stimulated with pitavastatin at a concentration of 1 micromolar for 4 hours.
|
| 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 RNAeasy clean-up columns (Qiagen). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc.
|
| |
|
| Hybridization protocol |
10 micrograms of fragmented cRNA were hybridised (45 degrees CelsiusC, 16 hours). Hybridization was controled by use of the GeneChip Eukaryotic Hybridization Control Kit (Affymetrix). Washing and staining was 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 |
KLF2 siRNA with 4h statin treatment
|
| Data processing |
Affymetrix Microarray Suite v5.0 with target intensity=100
|
| |
|
| Submission date |
Oct 07, 2011 |
| Last update date |
Jan 01, 2013 |
| Contact name |
Takahide Kohro |
| E-mail(s) |
kohro@lsbm.org
|
| Phone |
81-3-5452-5237
|
| Organization name |
LSBM, RCAST
|
| Street address |
|
| City |
4-6-1, Komaba, Meguroku |
| State/province |
Tokyo |
| ZIP/Postal code |
153-8904 |
| Country |
Japan |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE32693 |
Expression data of statin treated HUVEC cells transfected siRNA KLF2 or KLF4. |
|
