GSM8797344: High-Dose Tamoxifen (HDT) Group; Mus musculus; RNA-Seq
Gastric corpus tissues were harvested and dissociated into single-cell suspensions. Tissues were enzymatically digested using collagenase IV (2 mg/ml) and DNase I (40 U/ml) at 37°C for 30 minutes. Cells were then filtered, washed, and stained with viability dyes for live-cell sorting. Fluorescence-activated cell sorting (FACS) was performed using a BD FACSAria system, based on CD326+ and tdTomato+ expression for lineage tracing. Sorted cells were resuspended in PBS with 0.04% BSA for downstream processing. Single-cell RNA sequencing libraries were prepared using the 10x Genomics Chromium Next GEM Single Cell 3' v4 kit, following the manufacturer's protocol. Cells were encapsulated in a gel bead-in-emulsion (GEM) system, and poly-A RNA transcripts were reverse-transcribed into cDNA containing Illumina R1 primer sequences, Unique Molecular Identifiers (UMIs), and 10x barcodes. Post-amplification cDNA was purified using SPRIselect beads, and libraries were constructed with Illumina R2 sequencing adapters and sample indices for paired-end sequencing.