|
| Status |
Public on Feb 27, 2012 |
| Title |
HUVEC_Erg antisense_24hr_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
HUVEC treated with Erg antisense for 24 hours
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Umbilical vein endothelial cells
|
| Treatment protocol |
HUVEC were treated with either Erg-specific or control antisense (known as GeneBloc) for either 24 hr or 48 hr
|
| Growth protocol |
HUVEC were maintained in EGM-2 media
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cells using the Rneasy kit (Qiagen) according to the manufacturer's instructions
|
| Label |
Biotin
|
| Label protocol |
2ug of total RNA is converted to cDNA via first and second strand synthesis using the GeneChip® Expression 3’-Amplification One-Cycle cDNA Synthesis kit, in conjunction with the GeneChip® Eukaryotic PolyA RNA Control Kit. Biotin labelled cRNA is synthesised from the double-stranded cDNA using the GeneChip® Expression 3’-Amplification IVT Labelling Kit
|
| |
|
| Hybridization protocol |
15µg of fragmented cRNA was made up into a hybridisation cocktail in accordance with the Affymetrix technical manual corresponding to a 49 format (standard)/64 format array. The hybridisation cocktail was added to the appropriate array and hybridised for 16hrs at 45°C. The array is washed and stained on the GeneChip® fluidics station 450 using the appropriate fluidics script.
|
| Scan protocol |
The array was inserted into the Affymetrix autoloader carousel and scanned using the GeneChip® Scanner 3000.
|
| Description |
S0172F031
|
| Data processing |
Differential gene expression analysis was carried out using Rosetta Resolver Data Analysis System (version 7.1, Rosetta Biosoftware). Statistical analysis was performed using false discovery rate (FDR)–corrected P values, following Benjamin-Hochberg Multiple Test Correction.
|
| |
|
| Submission date |
Oct 14, 2011 |
| Last update date |
Feb 27, 2012 |
| Contact name |
Graeme Miles Birdsey |
| E-mail(s) |
[email protected]
|
| Phone |
+442075948633
|
| Organization name |
Imperial College London
|
| Department |
National Heart and Lung Institute
|
| Street address |
Hammersmith Hospital, Du Cane Road
|
| City |
London |
| ZIP/Postal code |
W12 0NN |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE32984 |
Gene expression profiling of Human Umbilical Vein Endothelial Cells (HUVEC) after treatment with Erg or control antisense (GeneBloc) |
|
