|
| Status |
Public on May 13, 2015 |
| Title |
EML1 infected with the TEL-AML1 ccs cloned into thepWPI lentiviral vector - rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
EML1 hematopoietic progenitor cell line
|
| Organism |
Mus musculus |
| Characteristics |
cell line: EML1
treatment: TEL-AML1 ccs cloned into thepWPI lentiviral vector
|
| Growth protocol |
EML1 cells were grown in IMDM (Iscove Modified Dulbecco’s Medium) supplemented with 20% heat-inactivated horse serum and 15% BHK/MKL cell-conditioned medium. For B-cell differentiation, OP9 stromal cells were plated in 6-well plates in Alpha Minimum Essential Medium (α-MEM) with 20% of fetal bovine serum. When they reached confluency, 50 000 EML1 cells were plated on top of the stromal cells in their specific culture medium plus supplements (IL7 10 ng/ml; FLT3L 80 ng/ml; β-mercaptoethanol, 50 μM).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRIzol Reagent (Gibco), followed by clean up on RNeasy mini/midi columns (RNeasy Mini/Midi Kit, Qiagen).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 ug total RNA The size and the accuracy of quantitation of targets were checked by agarose gel electrophoresis of 2ug aliquots, prior to and after fragmentation. After fragmentation, targets were diluted in hybridisation buffer at a concentration of 150ug/ml.
|
| |
|
| Hybridization protocol |
Following fragmentation, 12.5 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Mouse Gene 1.0 ST Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station.
|
| Scan protocol |
Images were scanned using an Affymetrix GeneArray Scanner, using default parameters.
|
| Description |
EML1 cell line: Bone marrow cells of male BDF1 mice infected with the retroviral vector LRARalpha403SN. They can be induced with IL-7 and bone marrow stromal cells to express characteristics of pre-pro-B lymphocytes.
|
| Data processing |
standard Affymetrix protocol
|
| |
|
| Submission date |
Jan 13, 2015 |
| Last update date |
May 14, 2015 |
| Contact name |
Myriam Alcalay |
| E-mail(s) |
myriam.alcalay@ieo.eu
|
| Organization name |
European Institute of Oncology
|
| Department |
Experimental Oncology
|
| Lab |
Functional Genomics
|
| Street address |
Via Adamello 16
|
| City |
Milan |
| ZIP/Postal code |
20139 |
| Country |
Italy |
| |
|
| Platform ID |
GPL11533 |
| Series (1) |
| GSE64919 |
Genes regulated in EML1 cells expressing the TEL-AML1 oncogene after 5 and 7 days of treatment with IL7 and FLT3 ligand. |
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