title: GSM8232701: Foxf2-ECKO, replicate 1, scRNA-seq; Mus musculus; RNA-Seq
library_name: GSM8232701
library_construction_protocol: Brain endothelial cells (BECs) were isolated from whole mouse brain as previously described (see methods in manucript) and sorted using Fluorescence-activated cell sorting (FACS). Following CD31 enrichment, cell suspension was resuspended in Flow Cytometry buffer (Invitrogen, Cat# 00-4222-26) and stained with CD11b (Invitrogen, Cat# 53-0112-82), CD45 (Invitrogen, Cat# 53-0451-82) and Fixable Viability Dye eFluor™ 780 (Invitrogen, Cat# 65-0865-14). Doublets and microglia population (defined as CD11b and CD45+) were gated out prior to FACS. Viable ECs were sorted into RPMI media (Invitrogen, Cat# 11835030) supplemented with 10% FBS. Fresh single cell suspensions were centrifuged and resuspended in PBS containing 0,04% ultra-pure BSA. Libraries for single-cell sequencing (scRNA-seq) were prepared using the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10X Genomics) following manufacturer's instructions.