|
| Status |
Public on Apr 30, 2012 |
| Title |
DN1-isotype controls-2 |
| Sample type |
RNA |
| |
|
| Source name |
DN1-isotype controls
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
tissue: thymus
|
| Treatment protocol |
C57BL/6 mice were injected with anti-DLL4 or isotype control mAb. Two weeks later, mice were sacrified and thymi were harvested. Total thymocytes were isolated from treatment and control animals. The rest of the thymocytes were stained and two cell subsets were sorted based on the expression of CD4 and CD8 cell surface markers: DN cells (CD4-CD8-) and DN1(CD4-CD8-CD44+CD25-).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Samples were transfered to TRIzol (Invitrogen). Total RNA was isolated according to manufacturer´s instructions.
|
| Label |
Cy3
|
| Label protocol |
RNA from each sample was amplified by TargetAmp 2-Round Aminoallyl-aRNA Amplification Kit (Epicentre). Cy3-CTP was incorporated into amplified cRNA from 500 ng of total RNA using QuickAmp RNA Amplification Kit (Agilent).
|
| |
|
| Hybridization protocol |
Cy3 labeled cRNA from each sample was then hybridized to a custom Agilent array comprising of 43538 60mer oligos covering mouse transcriptome. The hybridization and wash of the arrays were performed according to the Agilent protocol.
|
| Scan protocol |
Arrays were scanned on an Agilent Microarray scanner.
|
| Description |
DN1 (CD4-CD8- CD44+CD25-) cells sorted from total thymocytes treated with isotype control
|
| Data processing |
The data was extracted from scanned array images using Agilent Feature Extraction Software 9.5. Global normalization was done by normalizing each chip to the 75 percentile of the measurements taken from that chip.
|
| |
|
| Submission date |
Sep 23, 2010 |
| Last update date |
Apr 30, 2012 |
| Contact name |
Wen Zhang |
| Organization name |
Regeneron Pharma.
|
| Street address |
777 Old Saw Mill River Road
|
| City |
Tarrytown |
| ZIP/Postal code |
10591 |
| Country |
USA |
| |
|
| Platform ID |
GPL10964 |
| Series (1) |
| GSE24296 |
Molecular mechanism underline immune cell type population shift upon anti-DLL4 treatment |
|
