Ferret single cell transcriptomics analysis

We applied 10X Genomics scRNA-seq technology to analyze molecular signatures of neural progenitor subtypes at stages covering early-to-late neurogenesis and gliogenesis (E25, E34, E40, P1, P5, P10) in ferret (Mustela putorius furo). We isolated cell populations in two different ways to enrich progenitor subtypes ; (1) FACS-sorting of fluorescent cells from tissues that had been electroporated at E30 or E34, with AzamiGreen (AG) expression vector (Karasawa et. al. 2003) under the control of Hes5 promoter with a d2 degradation signal (Ohtsuka et al. 2006), (2) taking cells from the cerebral cortex after discarding the CP, thereby collecting cells from the VZ, SVZ and intermediate zone (IZ) with a less abundance of mature neurons in the CP. These two cell populations were combined for downstream analyses.