organism: Mus musculus
description: Protocols: Mouse aorta was first digested for 1 hour at 37°C in 1 mg/mL collagenase I (Gibco, Life Technologies), 1 mg/mL dispase (Gibco, Life Technologies), 150 µg/mL DNase-I (Sigma-Aldrich) before passing the suspension through a 70 µmol/L cell strainer. Lineage+ cells were then depleted from tumor cell suspensions via MACS cell separation according to the manufacturer's instructions (Miltenyi Biotech). Cell number and viability for each sample was assessed by using 0.4% Trypan blue solution and a hemocytometer. Single-cell suspensions were then used for flow sorting or analysis by flow cytometry. ENSG ids were converted to gene symbols using tximport. RNA was extracted from sorted endovascular progenitor (EVP), transit amplifying (TA), and definitive differentiated (D) cells using a QIAGEN mini kit (Qiagen) according to the manufacturer's instructions. RNA quality and concentration was assessed by using the Agilent 2100 Bioanalyser (Agilent Technologies). Five to 100 ng of RNA was used for cDNA synthesis using the Superscript III Reverse Transcription Kit (Invitrogen). Total isolated RNA of appropriate RNA integrity number (>4) was prepared using the TruSeq stranded total RNA library preparation kit as per the manufacturer's instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-03-31T17:01:55Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-08-14T13:33:54Z
attributes.tag: External Id
attributes.value: SAMEA4828309
attributes.tag: INSDC center name
attributes.value: Division of Genomics of Development and Disease, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, Australia, 4072
attributes.tag: INSDC first public
attributes.value: 2019-03-31T17:01:55Z
attributes.tag: INSDC last update
attributes.value: 2018-08-14T13:33:54Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7148:Sample 2
attributes.tag: age
attributes.value: 8 to 12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: aortic endothelial cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: organism part
attributes.value: aorta
attributes.tag: phenotype
attributes.value: CD31_hi/VEGFR2_hi
attributes.tag: sample name
attributes.value: E-MTAB-7148:Sample 2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: C57BL/6