RNA-seq of LUHMES Neurons
LUHMES passage 4 containing 5 million cells was flash frozen. RNA was isolated using Qiagen RNeasy Plus Mini Kit (Cat. No. 74134).  To capture non-coding RNAs, expression was studied after ribosomal RNA depletion. Strand-specific and dual-barcode indexed RNA-seq libraries were generated from 450 ng total RNA each using the Kapa RNA-seq Hyper kit (Kapa Biosystems-Roche, Basel, Switzerland) and both the QIAseq FastSelect5S/16S/23S ribodepletion and FastSelect rRNA Plant reagents (Qiagen, Hilden Germany) in combination, following the instructions of the manufacturers.  The fragment size distribution of the libraries was verified in an automated electrophoresis platform on the TapeStation (Agilent, Santa Clara, CA).   The libraries were quantified by fluorometry on a Qubit instrument (Life Technologies, Carlsbad, CA) and pooled in equimolar ratios. The pool was quantified by qPCR with a Kapa Library Quant kit (Kapa Biosystems) and sequenced on an Illumina NovaSeq 6000 (Illumina, San Diego, CA) with paired-end 150 bp reads.