|
| Status |
Public on May 06, 2019 |
| Title |
RAW264.7 cells_IR-61_repeat1 |
| Sample type |
RNA |
| |
|
| Source name |
RAW264.7 cells, IR-61, repeat1
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Abelson murine leukemia virus-induced tumor; ascites
cell line: RAW264.7
agent: IR-61
strain: BALB/c
gender: male
age: adult
|
| Treatment protocol |
RAW264.7 cells was treated with vehicle control or IR-61 for 24h in DMEM medium supplemented with 10% heat-inactivated fetal bovine serum. The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RAW264.7 cells were treated with vehicle control or IR-61 for 24 h and then RNA was extracted, reverse transcription and analyzed.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) 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 ( Agilent p/n G2565BA) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Gene expression after 24h in IR-61-treated RAW264.7 cells
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.0.0.1 (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.
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| |
|
| Submission date |
May 05, 2019 |
| Last update date |
May 06, 2019 |
| Contact name |
Yawei Wang |
| E-mail(s) |
1130483585@qq.com
|
| Organization name |
Third Millitary Medical University
|
| Street address |
30 gaotanyan center street
|
| City |
Chongqing |
| ZIP/Postal code |
400038 |
| Country |
China |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE130718 |
Determinaton the effect of IR-61 on macrophage gene expression |
|
