|
| Status |
Public on Jul 01, 2019 |
| Title |
EL2 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse Liver Tissue Treated with Small Extracellular Vesicles
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: Nude
tissue: Liver
treatment: SW620-derived Small Extracellular Vesicles
|
| Treatment protocol |
Control group was injected with PBS 10 times every other day in 21days.
Purified SW620-derived sEVs were injected to nude mice via tail vein every other day for 21 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted by mirVanaTM RNA Isolation Kit (Applied Biosystem p/n AM1556) following the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.2 μg RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
0.6 μg of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/μg cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 22.5μl containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers' instructions. On completion of the fragmentation reaction, 22.5μl of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent mouse Gene Expression（8*60K）for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 4x180k array slides (Scan Area 61x21.6 mm, Scan resolution 3um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
mRNA expression.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.1.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities as the raw data. Raw data were normalized in quantile algorithm with GeneSpring 13.0 (Agilent). Probes that had at least 1 out of 2 samples flagged as Detected were maintained.
|
| |
|
| Submission date |
Jun 23, 2017 |
| Last update date |
Jul 01, 2019 |
| Contact name |
yingkuan Shao |
| E-mail(s) |
ykshao@zju.edu.cn
|
| Organization name |
Zhejiang Universit
|
| Street address |
Jiefang Road 88
|
| City |
Hangzhou |
| ZIP/Postal code |
310000 |
| Country |
China |
| |
|
| Platform ID |
GPL21163 |
| Series (1) |
| GSE100435 |
Colorectal Cancer-derived Small Extracellular Vesicles Create an Inflammatory Pre-metastatic Niche through Macrophage Polarization in Liver Metastasis |
|
