Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Single cells from spinal cords were isolated for single-cell RNA-sequencing as described previously (Vanlandewijck et al. Nature 2018). Mice were perfused with ice-cold PBS and spinal cords were dissected. Briefly, after enzymatic digestion, mechanical dissociation and myelin debris removal, eluted cells were incubated with mouse Fc Blocker (clone 2.4G2, BD Pharmingen) stained with CD45-FITC (clone 30-F11, BioLegend), CD31-APC (clone MEC13.3, BD Pharmingen), and DAPI. Equal numbers of viable DAPI-CD45+ leukocytes and DAPI-CD31+ ECs were sorted with BD Influx Cell Sorter (BD Biosciences) into PBS supplemented with 0.04% BSA. Subsequent cDNA purification, amplification (12 cycles) and library construction (sample index PCR 14 cycles) was performed as instructed. Sample libraries were sequenced on the Illumina NovaSeq 6000 system using S1 flow cell (Illumina)