Indices 1 12

Libraries were prepared from total RNA using the NEBNext Ultra II Directional RNA-Seq Kit (New England BioLabs, Ipswich, MA, USA). mRNA was isolated using the Poly(A) RNA Selection Kit (Lexogen, Vienna, Austria). mRNA was used for cDNA synthesis and shearing following the manufacturer’s instructions. Indexing was performed using Illumina indices 1–12. Enrichment was carried out by PCR. Subsequently, libraries were checked using the Agilent 2100 Bioanalyzer and DNA High-Sensitivity Kit to evaluate the mean fragment size. Quantification was performed using the Library Quantification Kit and StepOne™ Real-Time PCR System (Life Technologies, Carlsbad, CA, USA). High-throughput sequencing was performed on paired-end reads (2 × 100 bp) using the HiSeq 2500 platform (Illumina, San Diego, CA, USA.).

Total RNA was isolated using TRIzol reagent (Invitrogen, 15596018), and RNA quality and quantification were performed using an Agilent 2100 bioanalyzer (Agilent Technologies) and an ND-2000 Spectrophotometer (Thermo). Library preparation was performed using the NEBNext Ultra II Directional RNA-Seq Kit (NEW ENGLAND BioLabs, E7760L). Using a Poly(A) RNA Selection Kit, mRNA was extracted (LEXOGEN, 157.96) and used according to the manufacturer’s instructions for cDNA synthesis and shearing. Indexing was performed using Illumina indices 1–12 and was enriched by PCR. The mean fragment sizes were evaluated, and libraries were checked using a TapeStation HS D1000 Screen Tape (Agilent Technologies). Quantification was performed using the StepOne Real-Time PCR System (Life Technologies), and high-throughput sequencing was performed using NovaSeq 6000 (Illumina). FastQC was used to control the quality of the raw sequencing data, and adapters and low-quality reads (

Total RNA was isolated using TRIzol reagent (Invitrogen). RNA quality was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies), and the RNA was quantified using an ND-2000 Spectrophotometer (Thermo). Libraries were prepared from the total RNA using the NEBNext Ultra II Directional RNA-Seq Kit (New England BioLabs). mRNA was isolated using the Poly(A) RNA Selection Kit (Lexogen) and was used for cDNA synthesis and shearing, following the manufacturer’s instructions. Indexing was performed using Illumina indices 1–12. The libraries were enriched using PCR. Fragment size was evaluated using the Agilent 2100 Bioanalyzer (DNA High Sensitivity Kit). Quantification was performed using a library quantification kit and a StepOne RT‒PCR System (Life Technologies). High-throughput sequencing was performed as paired-end 100 sequencing using a NovaSeq 6000 instrument (Illumina).