title: GSM8713915: plaque cells from M-LipaKI mice; Mus musculus; RNA-Seq
library_construction_protocol: The ascending aorta, aortic arch, and descending thoracic aorta were excised, cleaned of perivascular adipose tissue, and cut into small pieces to facilitate enzymatic digestion. The aortic tissues were digested into single cells in 1 mL RPMI medium containing 4 U/mL of Liberase TM (Sigma, 5401127001), 60 U/mL of hyaluronidase (Sigma, H3506), and 60 U/mL of DNase I (Worthington Biochemical Corporation, LS006333) at 37°C for 40 min. The enzymes were then neutralized by adding 4 mL of RPMI medium supplemented with 10% FBS. The obtained cell suspensions were filtered through 100 μm cell strainers and stained in 500 μL FACS buffer including DRAQ5 (BioLegend, 424101,1:1000) and DAPI (BioLegend, 422801, 1:3000) for 10 min on ice. Viable singlets (DRAQ5+DAPI-) were collected into DMEM/F12 medium (Cytiva SH30261.01) containing 10% FBS, and immediately submitted to the Single Cell Analysis Samples were prepared using the 10x Genomics Chromium Single Cell 3' Reagent Kits v2 (mouse samples) according to the manufacturer's instructions by Single Cell Analysis Core at the JP Sulzberger Columbia Genome Center.