Total RNA extraction was performed on iSNs using the PureLink™ RNA Mini Kit (Invitrogen) following the manufacturer's directions (n = 3 independent cell culture preparations) Resulting FASTQ files were aligned with the human genome (genome build GRCh38.109) using the nf-core/rnaseq pipeline (Ewels et al., 2020). Briefly, FASTQ files were pre-processed to infer strandedness (Salmon), trim reads (Trim Galore!), and remove genome/ribosomal contamination (BBSplit and SortMeRNA, respectively). Reads were then aligned to reference genome using STAR and Salmon (Dobin et al., 2013; Patro et al., 2017). Quality control of read alignment was conducted using MulitQC, qualimap, and FastQC.