GSM6373836: Aortic cells_3_HFD_ADT; Mus musculus; OTHER
Mice were anesthetized and blood was extracted using cardiac puncture. The whole aorta was collected after left ventricular perfusion with 10 mL of phosphate-buffered saline (PBS) and quickly transferred to cold PBS. To prepare a single-cell suspension, perivascular adipose tissue was removed and whole aortas from three mice were cut into ~1 mm pieces and digested with an enzyme solution consisting of 10mg/ml Collagenase II (Sigma) and 1mg/ml elastase (Worthington) for 15 min at 37 °C. The cell suspension was strained through a 40-μm filter and centrifuged at 500g x 5 minutes. The single cell libraries were prepared with Chromium Next GEM Single Cell 3' Kit v3.11 (10x Genomics) with Chromium Next GEM Chip G Single Cell Kit (10x Genomics) according to manufacturer's protocol with Chromium Controller (10x Genomics). Between 5,000 and 10,000 single cells were used for GEM generation. Then the cDNAs were prepared and separated by SPRI beads for cDNA library and protein library, and checked on the Agilent Bioanalyzer with High Sensitivity DNA kit (Agilent) Prepared cDNAs were used for library preparation for sequencing and final libraries were checked on the Agilent Bioanalyzer with DNA 1000 kit (Agilent). The libraries were sequenced with Illumina Nova-sequencer at a depth of 70,000 - 120,000 reads per cell.