RLPostC_rep2
On day 7 post-injury, a 6-mm segment of spinal cord tissue (3 mm centered on the lesion site) was harvested for single-cell RNA sequencing (scRNA-seq). Each group included four mice. The tissue was enzymatically dissociated using a cocktail of collagenase and DNase I, followed by filtration through a 40 m cell strainer to obtain a single-cell suspension. Cell viability was assessed using trypan blue staining and confirmed to be >85% prior to library preparation. Single-cell libraries were prepared using the 10x Genomics Chromium platform following the standard Chromium Next GEM Single Cell 3 Reagent Kits v3.1 protocol. The target cell recovery was 8,00010,000 cells per sample. Libraries were sequenced on an Illumina NovaSeq 6000 platform using 150 bp paired-end reads. Raw sequencing data were processed using Cell Ranger (v6.1.1) for quality control, alignment, and generation of gene-cell count matrices, using the mm15 reference genome.