Single-cell RNA sequecing of sensory ganglia meningeal leukocytes after neuropathic pain using 10X Genomics plataform

Sorted CD45⁺ meningeal immune cells from dorsal root ganglia (DRGs) were obtained as described in the section “Fluorescence-activated cell sorting (FACS) of DRG meningeal cells” and processed for droplet-based single-cell RNA sequencing using the 10x Genomics Chromium platform. Single-cell suspensions with viability >90% were loaded into the Chromium Controller, and libraries were generated using the Chromium Next GEM Single Cell 3′ Reagent Kits v3.1 according to the manufacturer’s instructions. Following encapsulation and barcoding, cDNA synthesis and amplification were performed, and library quality was assessed using a High Sensitivity D1000 ScreenTape Assay on an Agilent 4200 TapeStation. Libraries were sequenced on an Illumina HiSeq 4000 system.