RNA-seq of HEK293 cells overexpressing NOCT ?(2-15)-3F with controls overexpressing GST-3F

To investigated the effect of cytoplasmically localized human NOCT on transcriptome-wide RNA levels, we generated HEK293 cell lines that stably overexpress NOCT ?(2-15)-3F construct, which lacks the MTS and is localized to the cytoplasm. As a negative control, we also generated cells that stably express glutathione S-transferase-3x FLAG (GST-3F). Three independent clonal cell lines were generated for each construct, and served as biological replicates. Total RNA was isolated from each cell line, ribosomal RNA was depleted, and then stranded, whole transcriptome RNA sequencing was performed. Overall design: RNA-seq for each clonal cell line (3 lines expressing GST-3F and 3 lines expressing NOCT ?(2-15)-3F), by Illumina HiSeq 2500