characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-07-18T17:02:40Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-22T15:37:08Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104715459
characteristics.characteristic: External Id
characteristics.text: Laboratory of Angiogenesis and Vascular Metabolism VIB-KU Leuven Center for Cancer Biology (CCB)
characteristics.characteristic: INSDC center name
characteristics.text: 2018-07-18T17:02:40Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-22T15:37:08Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS2316842
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-6595:Sample 6
characteristics.characteristic: Submitter Id
characteristics.text: 10 to 12
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: endothelial cell
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000115
characteristics.characteristic: cell type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Mouse endothelial cells were isolated from perfused healthy lungs of CPT1A∆EC mice and their wild-type littermates. Briefly, before starting the perfusion, the mice were anesthetized with Ketamin/Xylasine (500μL/mouse of Ketamine 0.65% / Xylasine 0.05% diluted in saline solution) and once the withdrawal reflex was absent in pelvic limbs, the perfusion procedure was started. Mice were perfused with 5 ml of a water based perfusion buffer containing 1.7 M NaCl, 84 mM KCl, 120 mM Hepes and 1 mM NaOH followed by perfusion with 5 ml of a DMEM based digestion buffer containing 0.1% collagenase II, collagenase I, 2 mM CaCl2, 1% antibiotic/antimycotic, at a perfusion rate of 2 ml/min. Lungs were dissected and further dissociated using the gentleMACS dissociator system. Next, samples were centrifuged, resuspended in 5ml of PBS/0.5% BSA and applied to a pre-separation 70 μm filter. The filtered cells were washed twice with PBS/BSA buffer and endothelial cells were selected with CD31 MicroBeads (Milenyi), further stained with viability dye, CD102-APC and CD45-PeCy7 and the fraction of viable, CD102+ and CD45- cells obtained by FACS sorting was directly collected into Trizol reagent. RNA was extracted using TRIzol (Life Technologies). Starting from 1 µg total RNA, poly-adenylated fragments were isolated, reverse transcribed and converted into indexed sequencing libraries using the KAPA stranded mRNA-seq kit (Sopachem, Eke, Belgium) RNA libraries were created using the KAPA stranded mRNA-seq kit (Sopachem, Eke, Belgium) according to the manufacturers instructions.
characteristics.characteristic: description
characteristics.text: wild type genotype
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: lung
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0002048
characteristics.characteristic: organism part
characteristics.text: wild type
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: phenotype
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000384
characteristics.characteristic: sex
characteristics.text: C57BL/6NJ
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0007732
characteristics.characteristic: strain
characteristics.text: Sample 6
characteristics.characteristic: title