Disruption of ADNP-KDM1A-GTF2I complex drives neural differentiation imbalance in Helsmoortel-Van der Aa syndrome
Disruption of ADNP-KDM1A-GTF2I complex drives neural differentiation imbalance in Helsmoortel-Van der Aa syndrome
Total RNA was extracted from fresh pellets of iPSC or NCS using the RNeasy Mini Kit (Qiagen, 74104). The H9 human embryonic stem cell (hESC) derived neural stem cells (hNSCs) were purchased from Invitrogen (N7800-100). no dissection required The hNSCs were cultured on Geltrex-coated dishes (Geltrex LDEV-free reduced growth factor membrane matrix, Thermo Fisher Scientific, A1413202) in KnockOut DMEM/F12 (Invitrogen, 12660012) supplemented with 2 mM L-Glutamine (Thermo Fisher Scientific, 25030024), 20 ng/ml EGF (Peprotech, 315-09), 20 ng/ml FGF (Peprotech, 100-18B) and 2% of StemPro Neural Supplement (Thermo Fisher Scientific, A1050801). The media was refreshed every other day and cells were routinely passaged with Accutase (Sigma-Aldrich, A6964).  no treatment protocol was required nor performed on any cell line. no conversion protocol was required nor performed on any cell line. Total RNA was extracted from fresh pellets of iPSC or NCS using the RNeasy Mini Kit (Qiagen, 74104). Purified RNA was quantified using a NanoDrop spectrophotometer and RNA quality was checked with an Agilent 2100 Bioanalyzer using the RNA nano kit (Agilent, 5067-1512). RNA was isolated using the GenElute™ Mammalian Total RNA Miniprep Kit (Sigma-Aldrich, RTN350-1KT) Library preparation for RNA sequencing was performed according to TruSeq Total RNA sample preparation protocol (Illumina, RS-122-2202), starting from 250 ng - 1 μg of total RNA. cDNA library quality was assessed on Agilent 2100 Bioanalyzer, using the high sensitivity DNA kit (Agilent 5067-4626) Total RNA was prepared using the Illumina TruSEq Stranded mRNA library Prep kit