RNA-seq of Mus musculus: mNC tumors
Wild-type and Sox2 knockout (KO) tumors were dissected and immediately submerged in RNAlater (Thermo Fisher Scientific) at 4C for 24 hours, followed by storage at 80C until library preparation. Total RNA was extracted using the RNeasy Mini Kit (Qiagen) according to the manufacturers protocol. One microgram of RNA per sample was subjected to ribosomal RNA (rRNA) depletion and library preparation using the Illumina Stranded Total RNA Prep with Ribo-Zero Plus kit, following the manufacturers instructions. Library concentration and size distribution were assessed using the Agilent 4200 TapeStation. Sequencing was performed by St. Jude Childrens Research Hospital, generating approximately 50 million paired-end reads per sample