CT_in_vitro_1
total RNA was extracted from SVZ-derived neural stem cells cultured in the neurosphere assay. RNA was isolated using Qiagen kits (Toronto, ON, Canada). Prior to library construction, rigorous quality control steps were implemented to ensure RNA purity and integrity. This involved initial assessment of RNA purity using Nanodrop2000 (Thermo Scientific; Waltham, MA, USA), agarose gel electrophoresis for RNA integrity and potential contamination, and reconfirmation of RNA integrity using the Agilent 2100 Bioanalyzer (Santa Clara, California, USA). Subsequently, mRNA was purified and randomly fragmented to initiate cDNA synthesis. For library construction, cDNA fragments of 150-200bp length were purified to a concentration of 1.5ng/l using AMPure XP beads (Beckman Coulter, Beverly, USA). Assessment of library effective concentration and RNA quality were performed using Agilent 2100 Bioanalyzer and Qubit2.0 (Thermo Scientific; Waltham, MA, USA). Finally, the libraries underwent sequencing on the HiSeq platform (Illumina, San Diago, CA, USA) following the manufacturers protocols.