Four-week-old human cortical neurons without astrocytes or 14 DIV mouse cortical neurons were used for gene expression analyses. Total RNA was isolated using mirVana kit (Invitrogen) or TRIzol (Invitrogen) according to the manufacturer’s instructions. RNA-seq analysis data are from a previous study14 (link). For qPCR, a total of 1 μg RNA was used to synthesize cDNA with the SuperScript® III First-Strand Synthesis System (Invitrogen). Quantitative RT-PCR was then performed using SYBR green (Applied Biosystems) and the QuantStudio™ 6 Real-Time PCR System (Applied Biosystems) or the StepOnePlus™ Real-Time PCR System (Applied Biosystems). Expression levels for all genes were normalized to the housekeeping gene GAPDH and expressed relative to the relevant control samples. Primers used for this study are summarized in Supplementary Table 3.
Kim N.S., Wen Z., Liu J., Zhou Y., Guo Z., Xu C., Lin Y.T., Yoon K.J., Park J., Cho M., Kim M., Wang X., Yu H., Sakamuru S., Christian K.M., Hsu K.S., Xia M., Li W., Ross C.A., Margolis R.L., Lu X.Y., Song H, & Ming G.L. (2021). Pharmacological rescue in patient iPSC and mouse models with a rare DISC1 mutation. Nature Communications, 12, 1398.
Other organizations :
University of Pennsylvania, Emory University, The University of Texas Health Science Center at San Antonio, Johns Hopkins Medicine, Johns Hopkins University, National Center for Advancing Translational Sciences, National Institutes of Health, National Cheng Kung University, Shanghai Jiao Tong University, Augusta University
Four-week-old human cortical neurons without astrocytes
14 DIV mouse cortical neurons
dependent variables
Gene expression levels
control variables
Total RNA isolation using mirVana kit (Invitrogen) or TRIzol (Invitrogen)
CDNA synthesis using SuperScript® III First-Strand Synthesis System (Invitrogen)
Quantitative RT-PCR using SYBR green (Applied Biosystems) and QuantStudio™ 6 Real-Time PCR System (Applied Biosystems) or StepOnePlus™ Real-Time PCR System (Applied Biosystems)
Expression levels normalized to the housekeeping gene GAPDH
controls
Relevant control samples
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