Illumina NovaSeq 6000 sequencing; scRNAseq of femoral artery segments in the context of hind limb ischemia in mice with or without an endothelial cell-specific deletion of transcription factor Prdm16
scRNAseq of femoral artery segments in the context of hind limb ischemia in mice with or without an endothelial cell-specific deletion of transcription factor Prdm16
Cell preparation, scRNA library preparation and sequencing. EC-Prdm16-/- or EC-Prdm16+/+ littermates (n=4 each) were treated with tamoxifen at P1 to P4 and were subjected to right FA ligation at 8 weeks of age. Three days later, FA segments downstream of the ligation and the corresponding segment on the unligated side were quickly dissected out and collected on ice. To obtain sufficient amounts of cells for sequencing, for each genotype and for each side, the segments from all 4 mice were pooled.  Single cell suspensions were converted to barcoded Drop-seq libraries by using the Chromium Single Cell 3'  Library, Gel Bead & Multiplex Kit and Chip Kit (10X Genomics; Pleasanton, California, USA), aiming for an estimated 4,000 cells per library. 4 mouse single cell libraries were prepared with the Chromium Single Cell 3' V2 Chemistry Library Kit, Gel Bead & Multiplex Kit and Chip Kit from 10X Genomics.