Untreated-2
For messenger RNA-sequencing, the library was constructed using QuantSeq 3 mRNA-Seq Library Prep Kit (Lexogen, Wien, Austria) according to the manufacturers instructions. Briefly, each 500ng total RNA was prepared, an oligo-dT primer containing an Illumina-compatible sequence at its 5 end was hybridized to the RNA, and reverse transcription was conducted. After degradation of the RNA template, second-strand synthesis was initiated by a random primer with an Illumina-compatible linker sequence at its 5 end. The double-stranded library was purified using magnetic beads to remove all reaction components and amplified to add the complete adapter sequences required for cluster generation. The finished library was purified from PCR components, and then high-throughput sequencing was performed as single-end 75 sequencings using NextSeq 500 (Illumina, CA, USA).