GSM9417247: Aorta, APOE4; Mus musculus; RNA-Seq
Mice were anesthetized and then perfused with phosphate-buffered saline. The whole aorta from the aortic arch to the ileac bifurcation was isolated and trimmed of excess fat tissues surrounding the aorta. The entire aorta was minced into small pieces in phosphate-buffered saline containing 2% BSA, and then incubated for 1 hr at 37 °C in the same buffer containing 450 U/mL collagen-1, 120 U/mL collagen XI, 60 U/mL DNase 1, 60 U/mL hyaluronidase, 1.5 mM CaCl2, and 1.25 mg/mL elastase. The resulting cell suspension was filtered through 70 µm filters, and washed several times to obtain single cell suspension. Quality control was assessed by live/dead cell staining with trypan blue staining. Cell suspensions obtained from 6 APOE2, 5 APOE3, and 7 APOE4 mice were pooled according to APOE genotype for single-cell RNA-seq analysis. Each pooled single cell suspension was loaded onto a 10x Genomics Chromium instrument to generate single-cell gel beads in emulsion and preparation of a bar-coded cDNA library for single cell RNA-sequencing. Quality of the cDNA library was confirmed using an Agilent Bioanalyzer. Full length sequencing was performed on two paired-end 75-bp flow cells using the NovaSeq S4  platform. An overall sequencing depth of approximately 400 million read pairs per sample.