Single-cell RNA-seq of Mouse Aorta

Mouse infrarenal abdominal aortas were collected and digested in an enzymatic mix (DMEM containing 10 mg/ml collagenase type II (C6885, Sigma Aldrich) and 1 mg/ml elastase (LS002292, Worthington Biochemistry)) for 15 minutes at 37 °C^{8 (link)}. The tissue suspension was filtered with a 40 μm cell strainer, then centrifuged at 500 g for 5 min. Cells were resuspended with DMEM containing 10% FBS. Single-cell suspensions from 4 aortas per group were pooled together as one sample. 5000 cells per sample were loaded on a Chromium Controller (10x Genomics). The scRNA-seq libraries were constructed using the Chromium Single Cell 3’ v3 Reagent Kit according to the manufacturers guidelines (10x Genomics). cDNA libraries were uniquely sample indexed and pooled for sequencing. A MiSeq (Illumina) sequencing run was used to sample balance on a NovaSeq S1 flowcell (Illumina) using a 28×91bp sequencing reaction targeting >50,000 reads/cell.