|
| Status |
Public on Nov 05, 2018 |
| Title |
NICD-OE-HEC mLSEC biological rep3 |
| Sample type |
RNA |
| |
|
| Source name |
Notch intracellular domain overexpressing primary murine liver sinusoidal endothelial cells, biological replicate no 3
|
| Organism |
Mus musculus |
| Characteristics |
genotype: Mut
tissue: liver
cell type: sinusoidal endothelial cells
|
| Treatment protocol |
Livers were perfused in situ through the portal vein with a collagenase/amino acid/saccharide solution (Sigma-Aldrich), dissected, mechanically disrupted, pooled from 3 mice, digested for 25 minutes at 38 °C in collagenase/Gey’s balanced salt solution (Sigma-Aldrich), and filtered through a mesh. Nonparynchymal cells were separated by a 19.3 % Nycodenz gradient. Afterward, liver sinusoidal endothelial cells (LSEC) were isolated by Magnetic-activated cell sorting (MACS) using CD146 MicroBeads (Miltenyi) according to manufacturers’ instructions.
|
| Growth protocol |
Clec4g-Cre(tg/wt) mice were crossed with Rosa26-N1ICD-IRES-GFP mice to generate Clec4g-Cre(tg/wt);Rosa26-N1ICD-IRES-GFP mice (NICD-OE-HEC).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with innuPREP RNA Mini Kit (Analytik Jena).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol.
|
| |
|
| Hybridization protocol |
Hybridization (16h x 45°C) was processed according to the standard Affymetrix protocol.
|
| Scan protocol |
Affymetrix GeneArray Scanner3000
|
| Description |
Gene expression data from primary murine liver sinusoidal endothelial cells with enhanced Notch signalling.
|
| Data processing |
The data were analyzed with a commercial software called JMP Genomics, version 7, from SAS. Gene expression profiling was performed using arrays of mouse Mogene-2_0-type from Affymetrix. A Custom CDF Version 22 with Entrez based gene definitions was used to annotate the arrays. The Raw fluorescence intensity values were normalized applying quantile normalization, RMA background correction and Medianpolish Probeset Summary
|
| |
|
| Submission date |
Sep 12, 2018 |
| Last update date |
Nov 05, 2018 |
| Contact name |
Carsten Sticht |
| Organization name |
University Heidelberg
|
| Department |
ZMF
|
| Street address |
Theodor-Kutzer-Ufer
|
| City |
Mannheim |
| ZIP/Postal code |
68169 |
| Country |
Germany |
| |
|
| Platform ID |
GPL24557 |
| Series (1) |
| GSE119875 |
Expression data from Notch intracellular domain overexpressing primary murine liver sinusoidal endothelial cells |
|
