|
| Status |
Public on Sep 07, 2012 |
| Title |
MCF7-shContr-nutlin3a_4h-2 |
| Sample type |
RNA |
| |
|
| Source name |
MCF7 cell line
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MCF7
treatment: infected with shContr and then treated with 10 μM nutlin3a for 4h
|
| Treatment protocol |
Cell lines were treated with 10 μM nutlin3a for 8 or 4h before analysis. Knock-down of STAT3 was performed using lentivirus system. MCF7 cells were infected with shContr or shSTAT3 and selected using puromicin selectivity marker. Obtained stably depleted clones were treated with nutlin3a for 4h, harvested and subjected for microarrays.
|
| Growth protocol |
MCF7 is a human breast carcinoma line in 10% FCS DMEM media
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from cell pellets was isolated with Rneasy Mini Kit (Qiagen), following the manufacterer's instructon. A fraction was quantitated by spectrophotometric measure using NanoDrop. RNA integrity was evaluated in agarose gel electrophoresis.
|
| Label |
biotin
|
| Label protocol |
cRNA synthesis was made using 3`IVT Express kit (Affymetrix) following manufacterer's instructions and starting with 200ng of total RNA
|
| |
|
| Hybridization protocol |
Labeled cRNA was fragmented in fragmentation buffer (5x buffer:200 mM Tris-acetate (pH 8.1)/500nM KOAc/150 mM MgOAc) before hybridization on the array. Fragmented cRNA was hybridized to the nicroarrays in 90ul hybridization solution. GeneTitan Hybridization, Wash and Stain kit for 3`IVT arrays (Affymetrix) was used for preparing hybridization mixes and processing plate array in GeneTitan
|
| Scan protocol |
Human Genome U219 array plate was processed in GeneTitan according to manufacturer's instructions
|
| Description |
tumor cell line
|
| Data processing |
The data were analyzed with RMA method in R
|
| |
|
| Submission date |
Jun 23, 2011 |
| Last update date |
Sep 08, 2012 |
| Contact name |
Fedor Nikulenkov |
| E-mail(s) |
[email protected]
|
| Phone |
0707922237
|
| Fax |
0707922237
|
| Organization name |
Fedor Nikulenkov
|
| Street address |
Nobels vag 16
|
| City |
Stockholm |
| ZIP/Postal code |
17177 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL13667 |
| Series (1) |
| GSE30183 |
Expression profiling of MCF7 cells upon nutlin3a treatment |
|
