GSM9530644: Perk KO, ZsGreen1-, 0 week WD, GEX; Mus musculus; RNA-Seq
At the end of each treatment timepoint, 3 mice of each genotype was euthanized following IACUC guidelines and the vasculature was immediately perfused with ice-cold PBS containing  actinomycin D. Working quickly, the ascending aorta of each mouse was isolated and independently enzymatically digested to single-cell suspensions, hashed, and sorted to separate viable ZsGreen+ and ZsGreen-populations. Cells were then pooled by Perk genotype and ZsGreen expression in a manner that ensured that all mice in each pool  were equally sampled, then pelleted and counted. A final pooling of the ZsGreen+ and ZsGreen- populations of each genotype was completed immediately before input into the 10x Genomics Chromium system to ensure equal numbers of SMC-derived and non-SMC-derived cells could be detected by scRNA-seq. Libraries were prepared according to manufacturer's instructions (Single Cell 3' v3 protocol, 10X Genomics).