|
| Status |
Public on Aug 27, 2018 |
| Title |
YFP+ liver EC, biological replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
liver
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL\6N
cell type: YFP+ (bone marrow-derived endothelial cell)
|
| Treatment protocol |
Mice were euthanized and liver tissue was collected. Briefly, minced liver tissue was incubated in collagenase solution for 30 mins. Next, erythrocytes were lysed with ACK-lysis buffer. Afterward, the single cells were stained with an antibody cocktail (CD45, CD31, CD146) for 20 mins on ice. Liver endothelial cells (EC) were isolated by FACS using CD45- CD31+ CD146+ surface marker profile. Further, liver EC were sorted according to YFP expression into YFP+ EC and YFP- EC.
|
| Growth protocol |
8 wk old mice were irradiated with 9 Gy (2 x 4.5 Gy). After 2 h resting period, irradiated mice were transplanted with 5000 YFP-labeled LSK cells intravenously.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were isolated using Arcturus PicoPure RNA isolation kit according to the manufacturer described protocol.
|
| Label |
Biotin
|
| Label protocol |
The Affymetrix 3’ IVT Pico Reagent Kit is used to generate labeled single-stranded cRNA from input amounts of 100pg total RNA. Label: Biotin
|
| |
|
| Hybridization protocol |
(according to user manual Affymetrix 3’ IVT Pico Reagent Kit) 6,6 ug of fragmented and labeled ds-cDNA were hybridized for 17 hr at 45°C on Affymetrix Mouse Clariom S Arrays.
|
| Scan protocol |
Gene Expression Microarrays were scanned using the Affymetrix GeneChip® Scanner 3000 according to GeneChip® Expression Wash, Stain and Scan Manual for Cartridge Arrays
|
| Data processing |
All intensity data were quarantile normalized.
|
| |
|
| Submission date |
Jun 28, 2018 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Mahak Singhal |
| E-mail(s) |
m.singhal@dkfz.de
|
| Organization name |
German Cancer Research Center (DKFZ)
|
| Street address |
Im Neuenheimer Feld 280
|
| City |
Heidelberg |
| ZIP/Postal code |
69120 |
| Country |
Germany |
| |
|
| Platform ID |
GPL23038 |
| Series (1) |
| GSE116377 |
Expression data from FACS-isolated liver endothelial cells |
|
