Total RNA was extracted from cultured cells using TRIzol reagent (Thermo Fisher Scientific) according to the manufacturer's protocol. RNA concentration and purity were measured using a NanoDrop spectrophotometer, and RNA integrity was evaluated using an Agilent 2100 Bioanalyzer. Samples with RNA integrity number (RIN) ≥ 6.0 were used for library preparation. RNA-seq libraries were prepared using a poly(A) mRNA enrichment strategy. Polyadenylated mRNA was isolated from total RNA using oligo(dT) magnetic beads. Purified mRNA was fragmented and reverse transcribed using random hexamer primers to generate first-strand cDNA, followed by second-strand synthesis. The resulting cDNA fragments were end-repaired, A-tailed, and ligated to Illumina sequencing adapters. Libraries were amplified by PCR and purified prior to sequencing.