|
| Status |
Public on Jun 16, 2019 |
| Title |
control_monolayer_biological rep 1 |
| Sample type |
RNA |
| |
|
| Source name |
MDA-MB-231 Breast cancer cells control_monolayer
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MDA-MB-231 cell line
phenotype: monolayer
genotype: control
|
| Treatment protocol |
The tetPMLIV MDA-MB-231 cells cultured in monolayer or sphere conditions were treated with doxycycline (sigma) for 48h to induce PMLIV OE.
|
| Growth protocol |
The PMLIV inducible MDA-MB-231 clone was cultured in DMEM (Gibco), supplemented with 10% FBS and Gentamycin in 5% CO2 and at 37oC.MDA-MB-231 spheres were grown in DMEM-F12 1:1 (Gibco) medium containing B27 (1:50) (Gibco) , bFGF (20ng/ml) (Gibco) , EGF (20ng/ml) (Gibco) and 0.2% methylcellulose, in ultralow attachment plates.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from control and PMLIV OE MDA-MB-231 cells using TRIzol reagent according to the manufacturer’s protocol.
|
| Label |
Biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol (Expression Analysis Technical Manual, 2005-2009, Affymetrix).Briefly, double-stranded cDNA is synthesized from 10 μg total RNA isolated from MDA-MB-231 cells. An in vitro transcription (IVT) reaction is then done to produce biotin-labeled cRNA from the cDNA. The cRNA is fragmented before hybridization.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C GeneChip® Human Transcriptome Arrays 1.0. GeneChips were washed and stained in the Affymetrix Fluidics Station.
|
| Scan protocol |
GeneChips were scanned using GeneChip® Scanner 3000.
|
| Data processing |
The data were analyzed using Transcriptome Analysis Console Software according to Affymetrix' instructions.
|
| |
|
| Submission date |
Sep 06, 2018 |
| Last update date |
Jun 16, 2019 |
| Contact name |
Joseph Papamatheakis |
| E-mail(s) |
[email protected]
|
| Organization name |
Foundation for Research and Technology - Hellas
|
| Street address |
Nikolaou Plastira 100
|
| City |
Heraklion |
| State/province |
Crete |
| ZIP/Postal code |
GR-70013 |
| Country |
Greece |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE119583 |
ProMyelocytic Leukemia protein IV (PMLIV) represses breast cancer proliferation by modulating the FOXO3-FOXM1 axis |
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