|
| Status |
Public on May 20, 2016 |
| Title |
GSC-25 DS treated |
| Sample type |
RNA |
| |
|
| Source name |
GSC25 treated with DS7423 at 100 nM for 24 hours
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: GSC-25
cell type: Glioma initiating cells
|
| Treatment protocol |
Cells were treated with or without 100nM BEZ235, 100nM DS7423 for 24 hours.
|
| Growth protocol |
These GBM neurospheres were cultured in DMEM/F12 medium containing B27 supplement (Invitrogen, Carlsbad, CA), basic fibroblast growth factor, and epidermal growth factor (20 ng/ml each).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
We extracted total RNA at 24 hour after treatment with control, BEZ235, DS7423, from 3 independent experiments. RNA was harvested using RNeasy mini Kits (Qiagen), DNase I digestion, and RNeasy MinElute Cleanup Kit (Qiagen) following manufacturer’s protocols. We performed synthesis of complementary RNA from total RNA and hybridization/scanning of U133 plus 2.0 microarrays using GeneChip products (Affymetrix) as described in the GeneChip manual.
|
| Label |
biotin
|
| Label protocol |
Complementary RNA synthesis and labeling/hybridization/scanning of U133A 2.0 microarrays using GeneChip products (Affymetrix) was done as described in the GeneChip manual (see below fields) by Expression Analysis, Inc.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Human HGU133Av2 Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneArray Scanner 3000 by Expression Analysis, Inc. GeneChips were scanned using the Affymetrix 3000 7G Scanner.
|
| Data processing |
The CEL files were preprocessed with the aroma package (http://aroma-project.org/) using the CDF file from GPL5356 and quantile normalization.
|
| |
|
| Submission date |
Mar 16, 2016 |
| Last update date |
May 20, 2016 |
| Contact name |
Dimpy Koul |
| E-mail(s) |
dkoul@mdanderson.org
|
| Organization name |
The University of Texas MD Anderson Cancer Center
|
| Department |
Neuro-Oncology
|
| Street address |
1515 Holcombe Ave
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
| |
|
| Platform ID |
GPL5356 |
| Series (1) |
| GSE79316 |
Response to PI3K/mTOR dual inhibition in Glioma initiating cells |
|
