title: GSM8776322: Pdk4 KO scRNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: The mice were euthanized with isoflurane, followed by immediate perfusion with 20 mL cold phosphate-buffered saline (PBS). Isolated fresh aortae were opened longitudinally and cut into 3 mm fragments, followed by cold digestion for 3 h at 4°C. Digesting cocktail is composed of .5 mg/mL collagenase A (10103578001, Roche), 0.5 mg/mL elastase (LS002294, Worthington Biochemical), 10 mg/mL Bacillus licheniformis protease (P5380, Sigma-Aldrich), 10 mg/mL Dispase II (D4693, Sigma-Aldrich),  125 U/mL DNase I (DN25, Sigma-Aldrich)  and 0.2% BSA in DMEM. Digested aortae were spined at 350 g for 5 min at 4°C, washed twice with 1% BSA in PBS, and stained with viability dye on ice for 20 min (1:2000, Thermo Fisher, 65-0865-14). Suspended cells were sorted for viable cells using flow cytometry, and collected in 0.04% BSA-PBS. following Nextera XT Library Prep Kit