Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: The infrarenal abdominal aortas pooled from five mice in each group were digested with enzyme solution [450 U/ml collagenase type I (Gibco, cat# 17100-017), 125 U/ml collagenase type XI (Millipore Sigma, cat# C7657), 60 U/ml hyaluronidase type I-s (Millipore Sigma, cat# H3506), and 60 U/ml DNase I (Millipore Sigma, cat# DN25)] for 1.5 hours at 37oC. The cell suspension was strained through a 70 μm filter and washed thrice with PBS. The single cells were resuspended in opti-MEM with 10% FBS. The cell suspensions were loaded on a 10x Genomics Chromium instrument to generate single cell gel beads in emulsion (GEMs). Libraries were prepared using the Chromium Single cell 3' Reagent Kits v2 (CG00052): Chromium Single Cell 3' Library & Gel Bead Kit v2 (PN-120237), Chromium Single Cell 3' Chip Kit v2 (PN-120236) and Chromium i7 Multiplex Kit (PN-120262). Libraries were sequenced on an Illumina HiSeq 4000 using paired-end sequencing with approximately >90% sequencing saturation.