title: GSM8657255: Mouse mesenchyme, 5 days post SNL surgery, batch 2; Mus musculus; RNA-Seq
library_construction_protocol: At acute (4-6 days after partial sciatic nerve ligation, PSNL) and chronic (2 months after PSNL) timepoints, C57BL/6J wild-type mice were transcardially perfused with phosphate buffered saline, after which ipsilateral sciatic nerves were dissected and trimmed in F12 solution (Gibco, New York, NY, Cat# 21765-029) to either a 0.5 cm piece (for scRNA-seq and flow data in Figure 5A) or a 1 cm piece around the lesion site (all other experiments). In either case, the distal and proximal pieces were kept at the same length (0.25 or 0.5 cm each). The suture was removed at this point, if still present. A single cell solution was generated as described previously (Liang et al., 2020), after which cells were stained with the nuclear marker DRAQ5, the live-dead stain DAPI, PercCP-Cy5-5-conjugated myelin beads, BUV737-CD45 to stain immune cells and BV786-Ter119 to stain erythrocyte precursors. DRAQ5+DAPI-,Myelin-,CD45-,Ter119- singlets were sorted into 1.5ml tubes, pre-coated with FBS, containing 3 µl of 3% BSA in PBS buffer. Library preparation (Chromium 10X  single cell 3' version 3)  proceeded following manufacturer's instructions and was carried out by specialists in the NIHR Maudsley Biomedical Research Centre (BRC) Genomics & Biomarker Core Facility at King's College London. Sequencing was performed on an Illumina NextSeq system.