|
| Status |
Public on Jun 20, 2017 |
| Title |
Parental cell line, biological rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Breast cancer cell line
|
| Organism |
Homo sapiens |
| Characteristics |
drug resistance: Parental sensitive cell line
cell line: BT474
|
| Treatment protocol |
T-DM1 resistant clones were derived from parental cells by prolonged exposure to T-DM1 following an initial dose-response study of T-DM1 (0.1 nM - 100 nM) over 5 days. Resistant BT474 T-DM1R cell lines were obtained by single-cell cloning and continuously cultured in the presence of 5 nM T-DM1 for 3 months. Clones were routinely tested for resistance.
|
| Growth protocol |
Cell lines were cultured at 37°C in a humidified atmosphere in the presence of 5% CO2 and 95% air. Cells were grown in Dulbecco´s Modified Eagle´s Medium (DMEM) supplemented with 10% FBS, 100 mU/mL penicillin and 100 µg/mL streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 3.5 ug of cRNA were hybridized for 16 hr at 45C on Human Gene 2.0 ST Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
Gene expression data from parental cell line BT474
|
| Data processing |
Data were processed using the RMA algorithm implemented in the Affimetrix Expression Console (version 1.4.1; Affymetrix). Differentially expressed genes were identified using Transcriptome Analysis Console (TAC version 3.0; Affymetrix).
|
| |
|
| Submission date |
Jun 19, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Sara García-Alonso |
| E-mail(s) |
[email protected]
|
| Phone |
0034923294815
|
| Organization name |
Centro de Investigación del Cáncer
|
| Lab |
Lab 15
|
| Street address |
Campus Miguel de Unamuno s/n
|
| City |
Salamanca |
| State/province |
Salamanca |
| ZIP/Postal code |
37007 |
| Country |
Spain |
| |
|
| Platform ID |
GPL16686 |
| Series (1) |
| GSE100192 |
Expression data from T-DM1 resistant clones |
|
