GSM7057422: SNCA_IL10; Mus musculus; RNA-Seq
Nigral tissues were dissociated using Adult Brain Dissociation Kit and OctoMACS Separator (both from Miltenyi Biotec) following manufacturer's instructions. Collected cells were stained by Zombie Aqua Fixable Viability Kit (Biolegend) to label dead cells. Cells were sorted using a FACSAria Fusion sorter (BD) excluding cells labeled with anti-O4, anti-PSA-NCAM and anti-ACSA2 antibodies and selecting hematopoietic cells stained by anti-CD45 antibody. Cells were collected in PBS 0.04% BSA to be processed for sequencing. After cell counting, cells deriving from the 4 different animals for each experimental condition were pulled together and nuclei were extracted following 10x Genomics protocol for Nuclei Isolation from Embryonic Mouse Brain Single Cell RNA-seq. A total of 1 million cells for each experimental condition were used as a starting point for nuclei extraction. After nuclei counting single cell libraries were prepared using Chromium Next Gene Expression Reagent Bundle (10x Genomics, cod. FC51000285).  For each experimental group a unique RNA-seq library was obtained and sequenced on a Nova-seq6000 platform at San Raffaele hospital omics facility