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 5' Gene Expression workflow according to the manufacturer's protocol. Dissociated cells were loaded onto the Chromium instrument for partitioning into Gel Bead-In-Emulsions (GEMs), where RNA transcripts were captured and reverse transcribed to generate barcoded cDNA. Following GEM recovery, cDNA was amplified and sequencing libraries were constructed using the appropriate 10x Genomics Chromium 5' gene expression reagents. Libraries were sequenced on the Illumina NovaSeq 6000 platform using paired-end reads. Sequence data were processed and aligned using the Cell Ranger pipeline. No major deviations from the standard protocol were introduced unless otherwise documented in study records.