|
| Status |
Public on Aug 06, 2015 |
| Title |
VEGF-down3 |
| Sample type |
RNA |
| |
|
| Source name |
C166 cells transduced with lentiviral vector (MOI10) encoding GFP + shRNA that downregulates VEGF-A
|
| Organism |
Mus musculus |
| Characteristics |
cell line: C166
cell type: endothelial cell
|
| Treatment protocol |
Cells were at 20% confluency when transduced with lentiviral vector enconding GFP or additionally shRNA that either up- or downregulates VEGF-A expression. The fresh medium was changed 72h after transduction. Cells were collected for analysis at day 7.
|
| Growth protocol |
C166 cells (ATCC:CRL-2583) were cultured in DMEM containing 10% FBS, 100 units/ml penicillin and 100 ug/ml streptomycin. Cells were grown in a humidified 95% air/5% CO2 incubator at 37C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Standard TRI-reagent total-RNA isolation protocol
|
| Label |
Biotin
|
| Label protocol |
Gene chip 3´IVT express kit standard protocol
|
| |
|
| Hybridization protocol |
Following fragmentation, 12,5 ug of cRNA were hybridized for 16 hr at 45C on Mouse Genome 430 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using Affymetrix GeneChip scanner.
|
| Description |
>99% of cells expressing GFP and VEGF-A downregulated
|
| Data processing |
Data were background adjusted with GC-RMA, quantile normalized, and median polish summarized using Matlab R2012b.
|
| |
|
| Submission date |
May 29, 2013 |
| Last update date |
Aug 06, 2015 |
| Contact name |
Mikko Petri Turunen |
| E-mail(s) |
mikko.turunen@uef.fi
|
| Organization name |
University of Eastern Finland
|
| Department |
AIVI
|
| Street address |
Neulaniementie 2
|
| City |
Kuopio |
| ZIP/Postal code |
70210 |
| Country |
Finland |
| |
|
| Platform ID |
GPL16225 |
| Series (1) |
| GSE47444 |
Off-target effects of VEGF-A regulation by shRNAs |
|
