|
| Status |
Public on Mar 08, 2017 |
| Title |
cells were treated with TGF-β1 (5ng/ml) and NCB-0846 (3µM) |
| Sample type |
RNA |
| |
|
| Source name |
Cell line
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: A549
cell type: non-small cell lung cancer cells
treated with: TGF-β1 (5ng/ml) and NCB-0846 (3µM)
|
| Treatment protocol |
After serum starvation for 24 h, cells were treated with DMSO (control), TGF-β1 (5ng/ml) or co-treated with TGF-β1 and NCB-0846 (3µM) or NCB-970 (3µM) for 48 h.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were prepared with RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled miRNA was prepared from 100ng total RNA using the miRNA Complete Labeling and hyb kit (Agilent) according to the manufacturer's instructions.
|
| |
|
| Hybridization protocol |
The dyied Cy3-labelled miRNA was Resuspended in 18 μL of nuclease-free water, 4.5 μL of the 10X GE Blocking Agent and 22.5 μL of 2x Hi-RPM Hybridization Buffer was added following the manufacturer’s instructions. After the sample was incubated at 100°C for 5 minutes, and immediately transfer to an ice water bath for 5 minutes, and hybridized to Human miRNA Microarray kit 8x60K rel.21.0 (Agilent) for 20 hours at 55°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 5 minute at room temperature with GE Wash Buffer 1 (Agilent) and 5 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately
|
| Scan protocol |
Microarrays were scanned immediately after washing on the Agilent SureScan Microarray Scanner (G2600D) using one color scan setting for 8x15k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green PMT is set to XDR Hi 100% and XDR Lo 5%).
|
| Description |
846
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.5.1.1(Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. The GeneView files were generated using Agilent's Feature Extraction software version 11.5.1.1
|
| |
|
| Submission date |
Mar 07, 2017 |
| Last update date |
Mar 08, 2017 |
| Contact name |
Teppei Sugano |
| Organization name |
Graduate School of Medicine,
|
| Street address |
Sendagi, Bunkyo-ku
|
| City |
Tokyo |
| ZIP/Postal code |
113-8603 |
| Country |
Japan |
| |
|
| Platform ID |
GPL21575 |
| Series (1) |
| GSE95766 |
Suppression of epithelial mesenchymal transition by a novel Traf2- and Nck-interacting kinase inhibitory compound |
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