Single-cell RNA sequencing of DRG Meninges leukocytes after neuropathic pain using 10X Genomics platform.

Sorted CD45⁺ meningeal immune cells from dorsal root ganglia (DRGs) were obtained 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 using paired-end chemistry. Raw sequencing data were processed using Cell Ranger, and filtered gene–cell count matrices were used for downstream analysis in Seurat (v5.3.0) implemented in R (v4.4.1).