|
| Status |
Public on May 16, 2012 |
| Title |
PC12_NGF_12h |
| Sample type |
RNA |
| |
|
| Source name |
PC12, 12h, 100 ng/ml NGF
|
| Organism |
Rattus norvegicus |
| Characteristics |
cell line: PC12
cell type: pheochromocytoma
|
| Treatment protocol |
Cells seeded on PolyD-lysine-coated plates were treated with 100 ng/ml murine NGF-7S (Invitrogen) in DMEM containing 1% HS and 0.5% FBS for 0, 12, 24 and 48 hr.
|
| Growth protocol |
Rat pheochromocytoma PC12 cells were cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% horse serum (HS, Invitrogen, Carlsbad, CA, USA), 5% fetal bovine serum (FBS, MP Biomedicals, Irvine, CA, USA), 100 U/mL penicillin and 100 μg/mL streptomycin at 37ºC under a humidified atmosphere of 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA containing miRNA was extracted from cells using the miRNeasy Mini Kit (Qiagen, Hilden, Germany).
|
| Label |
Cy3
|
| Label protocol |
One hundred nanograms of total RNA was labeled using the Agilent miRNA Complete Labeling and Hybridization Kit (Agilent Technologies) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
The hybridization used miRNA Complete Labeling and Hyb Kit according to the manufacturer's instructions.
|
| Scan protocol |
After hybridization, the microarrays were washed according to the manufacturer’s protocol and scanned on an Agilent DNA Microarray Scanner with the Scan Control software (Agilent Technologies).
|
| Description |
miRNA expression after 12hr in NGF-treated PC12
|
| Data processing |
The resulting images were processed, and raw data were collected using Agilent’s Feature Extraction software. The gene expression data were analyzed using GeneSpring GX 11 (Agilent). The signal intensity for each probe was normalized by a percentile shift, in which each measurement was divided by the 75th percentile of all measurements in its array.
|
| |
|
| Submission date |
Sep 14, 2011 |
| Last update date |
May 16, 2012 |
| Contact name |
Toshio Kojima |
| Organization name |
Toyohashi University of Technology
|
| Department |
Health Care Center
|
| Street address |
1-1 Hibarigaoka Tenpaku-cho
|
| City |
Toyohashi |
| ZIP/Postal code |
441-8580 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10906 |
| Series (1) |
| GSE32122 |
MicroRNA expression profiling of NGF-treated PC12 cells revealed a critical role for miR-221 in neuronal differentiation |
|
