|
| Status |
Public on Aug 03, 2009 |
| Title |
A2780 DMSO/BIO treatment 1 rep 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
A2780 ovarian adenocarcinoma cells, treated with DMSO
|
| Organism |
Homo sapiens |
| Characteristics |
A2780 ovarian adenocarcinoma cells, treated with DMSO
|
| Treatment protocol |
21 day continual culture in the presence of DMSO or 2.5mM GSK3 inhibitor 6-bromoindirubin-3’-oxime (BIO) with replenishment weekly on days 1 and 4.
|
| Growth protocol |
A2780 ovarian adenocarcinoma cells were cultured in RPMI medium containing 10 % Fetal Bovine Serum and 1 % L-glutamine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were rinsed in PE, trypsanised and total RNA was extracted using Nucleospin II RNA extraction kit (Macherey-Nagel) as per manufacutor's instructions.
|
| Label |
Cy3
|
| Label protocol |
800 ng of total RNA was labelled with Cy3 (DMSO treated cells) or Cy5 (BIO treated cells) using the Low RNA Input Linear Amplification Kit (Agilent technologies) as per manufacturor's instructions.
|
| |
|
| Channel 2 |
| Source name |
A2780 ovarian adenocarcinoma cells, treated with 2.5mM GSK3 inhibitor 6-bromoindirubin-3’-oxime (BIO)
|
| Organism |
Homo sapiens |
| Characteristics |
A2780 ovarian adenocarcinoma cells, treated with 2.5mM GSK3 inhibitor 6-bromoindirubin-3’-oxime (BIO)
|
| Treatment protocol |
21 day continual culture in the presence of DMSO or 2.5mM GSK3 inhibitor 6-bromoindirubin-3’-oxime (BIO) with replenishment weekly on days 1 and 4.
|
| Growth protocol |
A2780 ovarian adenocarcinoma cells were cultured in RPMI medium containing 10 % Fetal Bovine Serum and 1 % L-glutamine.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were rinsed in PE, trypsanised and total RNA was extracted using Nucleospin II RNA extraction kit (Macherey-Nagel) as per manufacutor's instructions.
|
| Label |
Cy5
|
| Label protocol |
800 ng of total RNA was labelled with Cy3 (DMSO treated cells) or Cy5 (BIO treated cells) using the Low RNA Input Linear Amplification Kit (Agilent technologies) as per manufacturor's instructions.
|
| |
|
| |
| Hybridization protocol |
100 μl of purified labelled total RNA was hybridised to Agilent 4x44K whole human genome arrays for 17 hours at 65 °C rotating at 10 rpm as per manufacturor's instructions.
|
| Scan protocol |
Slides were scanned using DNA microarray scanner model G2539A (Agilent Technologies) and Agilent Scan Control Software. Spot intensity and identity information was extracted using Agilent Feature Extraction Software.
|
| Description |
n/a
|
| Data processing |
All data was analysed within Genespring GX version 7.3.1. Data was normalised using a Lowess intensity dependant normalisation.
|
| |
|
| Submission date |
Jan 23, 2009 |
| Last update date |
Aug 03, 2009 |
| Contact name |
W. Nicol Keith |
| E-mail(s) |
n.keith@beatson.gla.ac.uk
|
| Organization name |
The University of Glasgow
|
| Street address |
Center for Oncology and Applied Pharmacology, University of Glasgow, CRUK Beatson Laboratories, Garscube Estate, Switchback Road
|
| City |
Glasgow |
| State/province |
Lanarkshire |
| ZIP/Postal code |
G61 1BD |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE14532 |
GSK3 activates telomerase by regulating dynamic behaviour of multiple transcription factors converging on the hTERT gene |
|
