Developmental Transcriptomic Analysis of Mouse CNS

FB, MB, HB, and CSC portions of the E10.5 CNS were isolated for comparison with the TF gene expression patterns of the NSC lines. The E10.5 stage was chosen because it is the developmental stage when the posterior-dominant Wnt expression profile is lost (Takemoto et al., 2016 (link)) and when the basic TF expression pattern in the CNS domains is established yet neurogenesis is minimal (Gary et al., 2004 (link); Thompson et al., 2014 (link); Andrews et al., 2019 (link); Sagner and Briscoe, 2019 (link)). Two litters of E10.5 embryos of C57BL/6J mouse, with 5 and 6 embryos of the right stage, were dissected to isolate the embryonic portions containing FB, MB, HB, and the CSC (anterior to the forelimb bud). In PBS containing 10 μg/mL Dispase II (Wako) and 0.1% BSA, eyes, otic vesicles, and attached non-CNS tissues were removed. The separate CNS domains were pooled, and RNAs were extracted using TRIzol™ Plus RNA Purification Kit (ThermoFisher).