|
| Status |
Public on May 14, 2014 |
| Title |
Lung_18 |
| Sample type |
RNA |
| |
|
| Source name |
Endothelial cells from E13.5 mouse lung
|
| Organism |
Mus musculus |
| Characteristics |
tissue: Lung
sample number: 18
cell type: GFP positive cells
strain: Homozygous Tie2-GFP transgenic mice (Jackson laboratory, strain 003658)
|
| Treatment protocol |
E13.5 Tie2-GFP embryos were micro-dissected for cortex and lungs. Cortex tissue was carefully cleared of the meninges and choroid plexus.Each tissue was diced with a scalpel, and enzymatically dissociated with a standard collagenase based assay before FACS sorting.
|
| Growth protocol |
Homozygous Tie2-GFP transgenic mice (Jackson laboratory, strain 003658) were used for BBB transcriptional profiling. Pregnant mice were obtained following overnight mating (day of vaginal plug was defined as embryonic day 0.5).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was purified with Arcturus PicoPure RNA isolation kit (Applied biosystemsTM), followed by NuGEN Ovation V2 standard linear amplification and hybridization to Affymetrix Mouse Genome 430 2.0 Array.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the NuGEN Ovation RAN Amplification system V2 and FL-Ovation cDNA Biotin Module V2 kits from 15-30ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 3.75 ug of cDNA were hybridized for 16 hr at 45C on GeneChip Mouse Genome 430 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450).
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000).
|
| Description |
Tie2-GFP reporter mouse to purify endothelial cells from E13.5 mouse brain
|
| Data processing |
The data were analyzed with Robust Multiarray Analysis (RMA) from the Bioconductor affy package using default analysis settings. The corrected PM probes were normalized using quantile normalization and expression measure calculated using median polish. The arrayQualityMetrics package in Bioconductor was used for QC analysis and no outliers were identified.
|
| |
|
| Submission date |
Apr 14, 2014 |
| Last update date |
May 14, 2014 |
| Contact name |
Meeta Mistry |
| E-mail(s) |
[email protected]
|
| Organization name |
Harvard School of Public Health
|
| Street address |
655 Huntington Ave
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE56777 |
Expression data of E13.5 mouse Blood Brain Barrier and lung endothelial cells |
|
