Single-cell transcriptome libraries were generated using the 10x Genomics Chromium platform and sequenced on an Illumina NovaSeq 6000 to produce paired-end RNA sequencing data for downstream expression analysis.
Single-cell gene expression libraries were prepared using the 10x Genomics Chromium Single Cell 3' Gene Expression workflow following the manufacturer's protocol. Cell suspensions were loaded onto the Chromium instrument to generate Gel Bead-In-Emulsions (GEMs), in which polyadenylated RNA transcripts were captured and reverse transcribed to produce barcoded cDNA. After GEM recovery and cDNA amplification, sequencing libraries were generated according to the corresponding Chromium Single Cell Gene Expression reagent kit instructions. Libraries were sequenced on an Illumina NovaSeq 6000 using paired-end reads. Sequence data were processed and aligned using Cell Ranger. No major protocol deviations were introduced unless otherwise documented.