Bulk RNA-seq of endothelial cells isolated from sciatic nerve and spinal cord of SOD1-G93 and control mice
Bulk RNA-seq of endothelial cells isolated from sciatic nerve and spinal cord of SOD1-G93 and control mice
For each replicate, 6-7 sciatic nerves and 3-4 spinal cords were pooled by genotype and dissociated. Control (non-transgenic) P90 sciatic nerves (3 independent replicates), SOD1G93A P90 sciatic nerves (3 replicates), Control P120 sciatic nerves (3 independent replicates), SOD1G93A P120 sciatic nerves (3 replicates), Control P90 spinal cords (3 replicates), SOD1G93A P90 spinal cords (3 replicates), Control P120 spinal cords (4 replicates), SOD1G93A P120 sciatic nerves (4 replicates). An average of 20,000 ECs were purified by FACS for each replicats after staining with rat anti-CD31 APC (1:50; BD Pharmingen Cat#551262) and rat anti-CD45.2 V405 (1:250; BD Biosciences Cat# 560697).  ECs (CD31+/CD45−) were sorted with BD FACSAria Fusion flow cytometer into 400l of Trizol-LS (Thermo Fisher Scientific Cat # 10296010). After quantification and quality control with Qubit 3.0 Fluorometer (Thermo Fisher Scientific) and Agilent 2100 Bioanalyzer, RNA-seq libraries were generated using the SMART-Seq Ultra Low Input RNA Kit (Illumina) (single-end, 100bp read length).