Quantifying Neuronal Gene Expression

Primary neurons were transduced at DIV 8 with lentiviral vectors encoding CTF-V5 or PCDH19 shRNA and relative controls (empty cFUW vector and scramble, respectively). Total mRNA was extracted at DIV 12 using NucleoSpin RNA Kit (Macherey-Nagel) and cDNA was synthesized with iScript cDNA Synthesis Kit (Biorad) according to the manufacturer’s instructions. RT-PCR was performed according to standard procedures (Longaretti et al., 2020 (link)) by using RPSA as endogenous control and the primers in Table S9.% of LSD1 splicing isoforms evaluation was performed as in (Zibetti et al., 2010 (link)).
hiPSCs-derived neurons were infected at DIV 1 with lentiviral vectors encoding PCDH19 shRNA and a relative control (scramble).
At DIV 21 or 35, as indicated, total mRNA was extracted using NucleoSpin RNA Kit (Macherey-Nagel). cDNA was synthesized using iScript cDNA Synthesis Kit (Bio-Rad) according to the manufacturer’s instructions. To measure mRNA expression, the primers in Table S9 were used.