|
| Status |
Public on Mar 23, 2021 |
| Title |
HuH-7-M exosome |
| Sample type |
RNA |
| |
|
| Source name |
HuH-7-M derived exosome by ultracentrifugation
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HuH-7
cell type: hepatocellular carcinoma (HCC) cell line
|
| Treatment protocol |
After medium was replaced with advanced DMEM medium, exosome is cllected from supernatant. The supernatant was filtered with a 0.22-mm Stericup and ultracentrifuged at 110,000g for 70 min at 4 ℃. The pellets were resuspended in PBS after ultracentrifugation.
|
| Growth protocol |
HuH-7 was cultured and maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS) and 500 µg/ml penicillin streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA samples were extracted from cells using miRNesy mini kit (Qiagen).
|
| Label |
Cy5
|
| Label protocol |
Extracted total RNA was labeled with Cy5 using the 3D-Gene miRNA labeling kit (Toray, Kamakura, Japan)
|
| |
|
| Hybridization protocol |
Hybridized for 16 h at 32 C with rotary shake (250 rpm). Hybridization buffer and washing protocol was followed by the protocol supplied by TORAY Industries, Inc..
|
| Scan protocol |
3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
|
| Data processing |
The raw data of each spot was normalized by substitution with a mean intensity of the background signal determined by all blank spots’ signal intensities of 95% confidence intervals.
|
| |
|
| Submission date |
May 26, 2020 |
| Last update date |
Mar 23, 2021 |
| Contact name |
Satoshi Kondo |
| Organization name |
Toray Industries,Inc.
|
| Street address |
10-1 tebiro 6-chome
|
| City |
Kamakura |
| State/province |
Kanagawa |
| ZIP/Postal code |
248-8555 |
| Country |
Japan |
| |
|
| Platform ID |
GPL21263 |
| Series (1) |
| GSE151169 |
Investigation of mechanism using highly intrahepatic metastatic cell lines of HCC |
|
