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Status |
Public on Aug 06, 2015 |
Title |
VEGF-down2 |
Sample type |
RNA |
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Source name |
C166 cells transduced with lentiviral vector (MOI10) encoding GFP + shRNA that downregulates VEGF-A
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Organism |
Mus musculus |
Characteristics |
cell line: C166 cell type: endothelial cell
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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.
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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.
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Extracted molecule |
total RNA |
Extraction protocol |
Standard TRI-reagent total-RNA isolation protocol
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Label |
Biotin
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Label protocol |
Gene chip 3´IVT express kit standard protocol
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|
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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.
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Scan protocol |
GeneChips were scanned using Affymetrix GeneChip scanner.
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Description |
>99% of cells expressing GFP and VEGF-A downregulated
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Data processing |
Data were background adjusted with GC-RMA, quantile normalized, and median polish summarized using Matlab R2012b.
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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 |
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|
Platform ID |
GPL16225 |
Series (1) |
GSE47444 |
Off-target effects of VEGF-A regulation by shRNAs |
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