|
| Status |
Public on Jan 05, 2012 |
| Title |
HL60 leukemic cells, empty vector, replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
HL60 leukemic cells, empty vector
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HL60
cell type: leukemic
treatment: empty vector
|
| Treatment protocol |
HL60 cells infected with MSCV-based retroviral vectors containing ER-tagged empty vector (CTL), MycA or MycD were seeded at a density of 0.5M/mL in RPMI medium containing 200nM of 4-OHT. After 16 hours, 100nM of retinoic acid or EtOH (vehicle) were added to the medium for 4 hours. Cells were then harvested and RNA was extracted.
|
| Growth protocol |
HL60 cells were cultured at 37ºC and 5% of CO2 in RPMI medium supplemented with 10% of fetal bovine serum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using the Qiagen RNeasy Mini Kit including DNAse I digestion to avoid potential contaminations of DNA following the manufacturer's recommendations.
|
| Label |
Cy3
|
| Label protocol |
500 ng of total RNA was labeled using Agilent's QuickAmp Labeling Kit following manufacturer instructions. Briefly: mRNA was reverse transcribed in the presence of T7-oligo-dT primer to produce cDNA. cDNA was then in vitro transcribed with T7 RNA polymerase in the presence of Cy3-CTP to produce labeled cRNA.
|
| |
|
| Hybridization protocol |
The labeled cRNA was hybridized to the Agilent Whole Human Genome 4x44k 60-mer oligo microarray (G4112F) according to the manufacturer's protocol. After hibridization, the microarrays were washed.
|
| Scan protocol |
Slides were scanned on an Agilent G2565BA microarray scanner at Green channel XDR Hi 100% and XDR Lo 100% green PMT and 5 um resolution.
|
| Description |
O5_1_2
|
| Data processing |
Intensity data was extracted using the Feature Extraction software (Agilent).
|
| |
|
| Submission date |
Oct 15, 2010 |
| Last update date |
Jan 05, 2012 |
| Contact name |
Luciano Di Croce |
| E-mail(s) |
[email protected]
|
| Phone |
+34933160132
|
| Organization name |
Centre for Genomic Regulation
|
| Street address |
Dr. Aiguader, 88
|
| City |
Barcelona |
| ZIP/Postal code |
08003 |
| Country |
Spain |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE24731 |
Gene regulation by phosphomimetic Myc and retinoic acid |
|
