GSM8488581: E18.0 Mesentery_All unsorted cells; Mus musculus; RNA-Seq
For unsorted cells, single cell suspension was obtained by dissociating E18.0 embryo intestines and mesenteries for 30 minutes to 1 hour at 37 °C with Liberase DH (Roche). After multiple washes, the cells were resuspended in red blood cell lysis buffer (Sigma) for 3 min. A total of 40000 cells were loaded onto a microwell cartridge of BD Rhapsody Express system (BD Biosciences). For FACS-sorted blood endothelial cells, single cell suspension was obtained by dissociating E18.0 embryo intestines for 1 hour at 37 °C with Liberase DH (Roche). Cell suspension was stained with conjugated antibodies in FACS buffer for 30 min. GFP+ ECs (around 9000 cells) were complemented with GFP- ECs to reach a total of 40000 cells and loaded onto a microwell cartridge of BD Rhapsody Express system (BD Biosciences) Single-cell whole transcriptome analysis libraries were prepared according to the manufacturer's instructions using a BD Rhapsody WTA Reagent kit (BD Biosciences)