Bulk RNA sequencing of PGAM5-overexpressed SH-SY5Y cells and VCP mutated human spinal cord organoids

To investigate downstream signaling upon PGAM5 upregulation, SH-SY5Y cells were transfected with Flag-tagged PGAM5 or empty vector control. Forty-eight hours post-transfection, cells were harvested and total RNA was isolated using TRIzol reagent.In parallel, valosin-containing protein (VCP)-mutant induced pluripotent stem cells (iPSCs) were differentiated into human spinal cord organoids. Organoids were collected at 35 days post-induction for total RNA extraction.Polyadenylated mRNA was enriched using oligo(dT) magnetic beads and reverse-transcribed into cDNA. Sequencing libraries were prepared and subjected to 150 bp paired-end sequencing on the Illumina HiSeq 2500 platform.