Gene expression profile at single cell level of aortic cells from 3 adult R26-DreER;Ki67-CrexER;R26-GFP (ProTracer) 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 (v1.1). Sequencing was performed on Illumina Hiseq X Ten PE150 platform. Fastq files were processed by CellRanger (v.4.0.0) pipeline.