GSM6592928: SMC-Yap1KO, aorta, Saline; Mus musculus; RNA-Seq
Samples of ascending aorta were harvested from  mice (n=3) and were pooled in Hanks' Balanced Salt Solution (HBSS, #14175095, Thermo Fisher Scientific, Waltham, MA) with 10% fetal bovine serum. Extra-aortic tissues, such as periaortic fat, were removed, and the aortic tissues were cut into small pieces and incubated with enzyme cocktail in HBSS containing Ca2+/Mg2+ (#14025092, Thermo Fisher) for 60 minutes at 37 °C. The cell suspension was filtered through a 40-μm cell strainer (CLS431750-50EA, Sigma-Aldrich), centrifuged at 350 g for 5 minutes, and resuspended by using cold HBSS (#14175095) with fetal bovine serum (5% vol/vol). Cells were stained with 4′, 6-diamidino-2-phenylindole (DAPI) and were sorted to select viable cells (≥95% viability) by using flow cytometry (FACS Aria III, BD Biosciences, Franklin Lakes, NJ). Single-cell suspensions were dispensed separately onto the Chromium Controller (10x Genomics, Pleasanton, CA) with a target of 10,000 cells/sample. The scRNA-seq library was constructed by using the Chromium Single-Cell 3′ v3 Reagent Kit (10x Genomics). Cells were mixed with barcoded primer-linked gel beads. Each cell was barcoded with a unique index, and every transcript within a single cell was barcoded with a unique molecular identifier (UMI). cDNAs were pooled, truncated, and amplified to generate cDNA libraries that were sequenced using a Next Generation sequencer NovaSeq 6000 (Illumina, Inc., San Diego, CA) in a pair-end fashion to obtain more than 80,000 reads per cell.