Spatial transcriptomic libraries were generated using the 10x Genomics Visium platform and sequenced on an Illumina NovaSeq 6000 to produce paired-end RNA sequencing data for spatial gene expression analysis.
Spatial gene expression libraries were prepared using the 10x Genomics Visium Spatial Gene Expression workflow following the manufacturer's protocol. Tissue sections were mounted on Visium slides, processed according to standard histology and permeabilization conditions, and RNA released from the tissue was captured by spatially barcoded oligonucleotides on the slide surface. Reverse transcription, second-strand synthesis, cDNA amplification, and library construction were performed according to the corresponding Visium reagent kit instructions. Libraries were sequenced on an Illumina NovaSeq 6000 using paired-end reads. Sequence data were processed and aligned using Spaceranger. No major protocol deviations were introduced unless otherwise documented.