organism: Mus musculus
description: Protocols: Aortas were dissected and digested for 30 mins at 37°C in 1mg/ml collagenase I (Gibco, Life Technologies, NY, USA), 1mg/ml Dispase (Gibco, Life Technologies, NY, USA), 150μg/ml DNase-I (Sigma-Aldrich, St Louis, MO, USA) before passing the suspension through a 70μM cell strainer. Dissociated single cells in PBS/BSA/EDTA were then incubated with various antibody combinations for multi-parameter flow acquisition and analysis. The following combinations of antibodies were used to assess the endothelial hierarchy populations: Rat anti-mouse VEGFR2 PE, CD31 PE-Cy7 and CD34 Alexa647 (Becton Dickinson, NJ, USA), Rat anti-mouse Lineage cocktail BV450 (Biolegend), Rat anti-mouse CD144 FITC (eBioscience). FACS was performed on a FACSAria cell sorter (Becton Dickinson, Franklin Lakes, NJ, USA). scRNA-seq was performed in triplicate using wild-type C57/BL6 mice. The 10X Genomics Chromium instrument (10X Genomics) was used to partition viable cells with barcoded beads, and cDNA from each cell was prepared using the Single Cell 3' Library, Gel Bead and Multiplex Kit (v2; 10X Genomics) as per the manufacturer's instructions. Library construction performed as per 10X Genomic Single Cell 3' RNA-seq protocol version 2
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-03-31T17:01:55Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-08-16T11:22:52Z
attributes.tag: External Id
attributes.value: SAMEA4832938
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: 2018-08-16T11:22:52Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7149:Sample 2
attributes.tag: age
attributes.value: 6 to 8
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: replicate
attributes.value: 2
attributes.tag: sample name
attributes.value: E-MTAB-7149:Sample 2
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: C57BL/6