Gene expression profile at single cell level of aortic cells from 2 adult R26-DreER;Ki67-CrexER;R26-GFP;Kdr rox/rox (Kdr-KO) mice after 8weeks of tamoxifen treatment.

The heterogeneity of endothelial cell proliferation is not addressed. We used single cell RNA sequencing (scRNA-seq) to analyze the diversity of aortic endothelial cell proliferation. Overall design: Aortic cells were isolated by digestion with Collagenase type 1 at 37? for 30 minutes. The cell suspension was filtered through a 70 µm cell strainer and then pelleted by centrifugation at 300g for 5 minutes. After washing cells with 500ml HBSS, cells were loaded onto the 10x Chromium controller the generate Gel Bead-in-Emulsions (GEMs), which were further processed into single cell 5' gene expression libraries using Chromium Single Cell 5' Library Kit (v2). Sequencing was performed on Illumina Nova6000 PE150 platform. Fastq files were processed by CellRanger (v.6.1.1) pipeline.