Total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. RNA purity was assessed by NanoDrop ND-1000 spectrophotometer, and integrity was evaluated with Agilent Bioanalyzer 2100 (RIN > 7.0). Poly(A) RNA was purified from 1 µg total RNA using Dynabeads Oligo(dT)25 (Thermo Fisher), fragmented with Magnesium RNA Fragmentation Module (NEB) at 94 °C for 5–7 min, and reverse-transcribed with SuperScript II (Invitrogen). Second-strand cDNA synthesis incorporated dUTP, followed by adapter ligation and size selection (~300 ± 50 bp) with AMPure XP beads. Libraries were amplified by PCR (8 cycles) and sequenced on an Illumina NovaSeq 6000 platform (LC-Bio Technology Co., Ltd., Hangzhou, China) with 2×150 bp paired-end reads.