|
| Status |
Public on Dec 15, 2018 |
| Title |
Triple stimuli + Rapamycin |
| Sample type |
RNA |
| |
|
| Source name |
mProx24 cell
|
| Organism |
Mus musculus |
| Characteristics |
cell type: proximal tubular cell
cell line: mProx24 cell
strain: C57B6
|
| Treatment protocol |
Subconfluent cells were incubated at 37°C and 1% O2 with same medium for 12 h, followed by stimulation with 150 μM of palmitate bound to bovine serum albumin (0.15%) for 9 hours. 0.4μg/ml Rapamycin was treated for 12 h before stimulation with palmitate.
|
| Growth protocol |
mProx24 cell were cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal calf serum, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37°C and 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
100 ng of total RNA were dephosphorylated with CIP at 37°C for 30 min, then ligated at 16°C for 2 h in the presence of T4 RNA ligase and 3μL pCp-Cy3.
|
| |
|
| Hybridization protocol |
Labeled samples were incubated at 55℃ for 20 hours with Hybri Spike In, Blocking Agent and HI-PRM Hybridazation Buffer.
|
| Scan protocol |
Images were scaned using Agilent Technologies Microarray Scanner, and quantified using Agilent Feature Extraction 10.7.3.1.
|
| Description |
1% HG Pal R
Triple stimuli + Rapamycin
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3.1 (Agilent) using default parameters (protocol GE1-v1_91 and Grid: 012391_D_20060331) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Dec 15, 2015 |
| Last update date |
Dec 15, 2018 |
| Contact name |
Shinji Kume |
| E-mail(s) |
[email protected]
|
| Phone |
(+81)77-548-2111
|
| Organization name |
Shiga University of Medical Science
|
| Department |
Medicine
|
| Street address |
Seta Tsukinowa-cho
|
| City |
Otsu |
| State/province |
Shiga |
| ZIP/Postal code |
520-2192 |
| Country |
Japan |
| |
|
| Platform ID |
GPL17912 |
| Series (2) |
| GSE76044 |
mProx24 cell: control vs diabetic condition (microRNA) |
| GSE76046 |
mProx24 cell: control vs diabetic condition |
|
