JMA_1
RNA Sequencing: RNA quality was first assessed using the Fragment Analyzer system (Agilent Technologies) with the High Sensitivity RNA kit. Only RNA samples with RNA Integrity Number (RIN)  7 were selected for library preparation. RNA quantification was performed using the Qubit RNA High Sensitivity Assay Kit (Thermo Fisher Scientific).. Libraries were prepared using the TruSeq Stranded mRNA Library Prep Kit (Illumina Inc.), following the manufacturer's instructions. mRNA was enriched using oligo(dT) magnetic beads to select for polyadenylated transcripts. The resulting libraries were evaluated on the Fragment Analyzer using the High Sensitivity NGS Fragment Analysis kit to verify fragment size distribution and library quality and quantified with the Qubit dsDNA High Sensitivity Assay Kit. Libraries were pooled equimolarly and denatured according to Illumina guidelines. The final pool was loaded onto a NextSeq 550 instrument (Illumina Inc., San Diego, CA) at a concentration of 1.5 pM, including 1% PhiX as an internal control. Sequencing was performed using the NextSeq 500/550 Mid-Output kit (300 cycles) in Paired-End mode(2150bp).