RNA-Seq of Rattus norvegicus
Total RNA was isolated and purified from the samples using TRIzol (Invitrogen, CA, USA). Subsequently, the quantity and purity of the total RNA were assessed using NanoDrop ND-1000 (NanoDrop, Wilmington, DE, USA) for quality control (QC). The mRNA containing PolyA tails was selectively captured using oligo(dT) beads (Dynabeads Oligo (dT), 25-61005, Thermo Fisher, USA). Following fragmentation of the captured mRNA under high-temperature conditions, cDNA synthesis was carried out using reverse transcriptase (1896649, USA). Moreover, double-stranded DNA was synthesized from the DNA/RNA hybrids using E. coli DNA polymerase I (NEB, m0209, USA) and RNase H (NEB, m0297, USA), with the incorporation of dUTP Solution (Thermo Fisher, R0133, USA) to generate blunt ends. An A base was then added to each end of the double-stranded DNA to facilitate the connection with adaptors containing a T base at the end, followed by size selection and purification using magnetic beads. The double-stranded DNA was digested using UDG enzyme (NEB, part number m0280, MA, US), and PCR was performed to generate a library with fragment sizes of approximately 300bp  50bp. Finally, the library was subjected to paired-end sequencing using the illumina Novaseq 6000 (LC Bio Technology CO.,Ltd. Hangzhou, China) following standard procedures, with a sequencing mode of PE150.