|
| Status |
Public on Apr 02, 2022 |
| Title |
Lymphatic endothelial cells_MSU_24 hour_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
Lymphatic endothelial cells, MSU, 24 hour
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Lymphatic endothelial cells
treatment: MSU for 24h
|
| Treatment protocol |
Lymphatic endothelial cells were seeded in 6-well (15*10^4 cells/well) for 24 h at 37°C. Then the cells were treated with PBS and 300μg/ml MSU for 24h, respectively.
|
| Growth protocol |
Lymphatic endothelial cells were incubated with growth medium comprising FBS (10%), streptomycin (100 μg/ml) and penicillin (100 U/ml) in a 5% CO2-humidified incubator. The cells was cultured at 37 °C with DMEM culture media.
|
| 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.
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| |
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| 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 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 |
MSU1
Gene expression after 24hours in MSU (300μg/ml) treated lymphatic endothelial cells.
Cell line
|
| 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). Probe that at least 1 out of 2 samples flagged as Detected were maintained.
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| |
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| Submission date |
Apr 01, 2022 |
| Last update date |
Apr 02, 2022 |
| Contact name |
Junli Chang |
| E-mail(s) |
[email protected]
|
| Organization name |
Long hua Hospital
|
| Street address |
Wanping South Road 725
|
| City |
shanghai |
| ZIP/Postal code |
200032 |
| Country |
China |
| |
|
| Platform ID |
GPL21163 |
| Series (1) |
| GSE199950 |
Lymphatic vessels drain monosodium urate and activate immunity in gouty arthritis |
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