|
| Status |
Public on Sep 20, 2012 |
| Title |
Control (mut-Elf5) infected MDA-MB-231 cell, replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
mut-2
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: MDA-MB231
tissue: breast cancer
|
| Treatment protocol |
MDA-MB231cells were infected with lentivirus containing either wild type or mutant Elf5 (in pLEX vector), and selected for 10 days with puromycin.
|
| Growth protocol |
Standard cloning process was used to clone coding sequence of Elf5 or DNA binding domain mutant Elf5 in pLEX vector. Lentivirus were produced in 293T cells.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were extracted using RNAeasy mini-prep kit from Qiagen
|
| Label |
Cy5
|
| Label protocol |
The testing RNA samples and universal human reference RNA (Strategene) were labeled with CTP-cy5 and CTP-cy3, respectively, with Agilent Quick Amp Labeling Kit.
|
| |
|
| Channel 2 |
| Source name |
Human universal reference RNA (Strategene)
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: mixed
|
| Treatment protocol |
MDA-MB231cells were infected with lentivirus containing either wild type or mutant Elf5 (in pLEX vector), and selected for 10 days with puromycin.
|
| Growth protocol |
Standard cloning process was used to clone coding sequence of Elf5 or DNA binding domain mutant Elf5 in pLEX vector. Lentivirus were produced in 293T cells.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were extracted using RNAeasy mini-prep kit from Qiagen
|
| Label |
Cy3
|
| Label protocol |
The testing RNA samples and universal human reference RNA (Strategene) were labeled with CTP-cy5 and CTP-cy3, respectively, with Agilent Quick Amp Labeling Kit.
|
| |
|
| |
| Hybridization protocol |
Each cy5 labeled testing sample was mixed with equal amount cy3-labeled reference sample and fragmented and blocked (Agilent Gene Expression Hybridization Kit), and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequential
|
| Scan protocol |
Scanned on an Agilent G2565BA scanner.
Images were quantified using Agilent Feature Extraction Software (version 9.5.3.1).
|
| Description |
Biological replicate 2, mutant Elf5 infected MDA-MB231 cells
|
| Data processing |
Agilent Feature Extraction Software (v 9.5.3.1) was used for background subtraction and linear Lowess normalization.
|
| |
|
| Submission date |
Sep 15, 2011 |
| Last update date |
Feb 29, 2024 |
| Contact name |
Yong Wei |
| E-mail(s) |
[email protected]
|
| Organization name |
Princeton University
|
| Department |
Molecular Biology
|
| Lab |
Kang
|
| Street address |
LTL227 Wathington Road
|
| City |
Plainsboro |
| State/province |
NJ |
| ZIP/Postal code |
08536 |
| Country |
USA |
| |
|
| Platform ID |
GPL4133 |
| Series (2) |
| GSE32144 |
MDA-MB-231 cell: infected with lentivirus to stably express mut-Elf5 vs WT-Elf5 |
| GSE32150 |
Elf5 inhibits epithelial mesenchymal transition in development and cancer metastasis through transcriptional repression of Snail2 |
|
