|
| Status |
Public on Aug 31, 2019 |
| Title |
EGF_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Hela cells cultured with EGF for a week and transfected pT-Rex-DEST31 vector contained full-length PAX1 cDNA, which was not inducing to expression .
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HeLa
|
| Treatment protocol |
PAX1 (NM_006192) was constructed by inserting a full-length cDNA product into a pT-Rex-DEST31 vector. The construct was then transfected into HeLa cells with Lipofectamine 2000 in Opti-MEM I reduced-serum medium at 37 °C in a 5% CO2 atmosphere for 4–5 hours, after which the medium was removed and replaced with fresh culture medium. Cells were selected by the antibiotic (G418) after 2 days in culture. The stablelly transfected cells treated EGF (50ng/ml). The PAX1 expression was induced by using doxycycline.
|
| Growth protocol |
Hela cells were cultured in Dulbecco's modified Eagle's medium (DMEM) or RPMI 1640 medium containing 10% (w/v) fetal calf serum, penicillin at 100 U/ml, streptomycin at 100 g/ml, and L-glutamine at 2 mmol/l.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The total RNA was harvested by using a Qiagen RNeasy kit. An additional DNase I digestion procedure was included in the isolation of RNA to remove contaminating DNA according to the manufacturer's protocol. The mRNA quality has achieved the ratio of 28S/18S >1 and RNA quality indicator (RQI) > 7 by using Agilent 2100 620 Bioanalyzer to send to the core service unit (Health GeneTech Corp., Taoyuan, Taiwan) for whole genome expression analysis.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays.
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Description |
control
SAMPLE 2
|
| Data processing |
The data were normalised using quantile normalisation with IlluminaGUI in R
|
| |
|
| Submission date |
Aug 23, 2017 |
| Last update date |
Aug 31, 2019 |
| Contact name |
Hung-Cheng Lai |
| E-mail(s) |
[email protected]
|
| Organization name |
Taipei Medical University
|
| Department |
Department of Obstetrics and Gynecology, School of Medicine, College of Medicine,
|
| Street address |
No.250, Wuxing St.
|
| City |
Taipei |
| ZIP/Postal code |
11031 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL22366 |
| Series (1) |
| GSE102986 |
PAX1-SET1B Interaction Drives Multiple Phosphatases and Inhibits Oncogenic Cascades |
|
