Illumina HiSeq 2500 paired end sequencing; RNA-seq of rat NSCs stimulated with lithium and/or photon radiation.
RNA-seq of rat NSCs stimulated with lithium and/or photon radiation.
NSCs were isolated from the cerebral cortices of day E15.5 embryos obtained from pregnant Sprague Dawley rats largely as described previously (S. Ilkhanizadeh et al.,Biomaterials, 2007 and T. Andersson et al., Mol Cell Neurosci, 2011). Cultured Neural Stem Progenitor Cells were collected from culture plates.  Cells were kept in enriched N-2 medium with 10 ng/ml of basic fibroblast growth factor (R&D systems, Minneapolis, MN, USA) added every 24 h and medium changed every alternate day to keep the cells in an undifferentiated and proliferative state. Cells were passaged every 5 days by detaching through scraping in HBSS. Thereafter, the cells were gently mixed in N-2 medium and plated at 1:4 density. To investigate LiCl (Sigma Aldrich, St. Louis, USA) effects, we exposed passage 3 (P3) NSPCs from 12 h before irradiation to LiCl (3 mM), as previously described.  A photon 60Co irradiation source was used to expose the NSPCs at a set distance of 80 cm and an absorbed dose of 2.5 Gy.  P3 cells were harvested 24 hours after irradiation for gene expression analysis. mRNA was extracted using RNeasy Mini Kit (Qiagen). Illumina TruSeq Stranded mRNA sample preparation kit with 96 dual indexes (Illumina, CA, USA)