|
Status |
Public on Sep 25, 2015 |
Title |
MOLM14_CA_24hrs_rep2 |
Sample type |
RNA |
|
|
Source name |
MOLM14 cells, CA, 24hrs
|
Organism |
Homo sapiens |
Characteristics |
cell line: MOLM-14 treatment: 10nM CA for 24hrs cell type: MLL-AF9-rearranged AML
|
Treatment protocol |
Cells were plated (12-well) in triplicate at 500,000 to 800,000 cells/mL and incubated in the presence of DMSO or 10nM CA for 24hrs. Cells were then washed twice with cold PBS, and snap frozen.
|
Growth protocol |
MOLM-14 cells were grown in RPMI-1640 supplemented with 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was isolated (RNeasy Plus Microkit, Qiagen or TRIzol, Life Technologies), processed, and hybridized to the Human U133 Plus 2.0 microarray (Affymetrix).
|
Label |
biotin
|
Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol using the GeneChip 3' IVT Express Kit from 100 ng total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
|
|
Hybridization protocol |
Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome U133 Plus 2 arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 7G Plus.
|
Description |
500,000-8000,00 cells were treated with 10nM CA for 24hrs
|
Data processing |
Arrays were corrected for background, normalized and log2-transformed using the rma function of the affy Bioconductor package.
|
|
|
Submission date |
Jan 15, 2015 |
Last update date |
Sep 25, 2015 |
Contact name |
Matthew D. Shair |
Organization name |
Harvard University
|
Department |
Chemistry and Chemical Biology
|
Lab |
Shair
|
Street address |
12 Oxford Street
|
City |
Cambridge |
State/province |
MA |
ZIP/Postal code |
02138 |
Country |
USA |
|
|
Platform ID |
GPL570 |
Series (2) |
GSE65014 |
Effect in MOLM-14 cells of 24hr cortistatin A treatment on gene expression |
GSE65161 |
Mediator kinase inhibition further activates super-enhancer-associated genes in AML |
|