Transcriptional response to Notch activation in SVG-A cells

SVG-A cells are immortalized fetal astrocytes that respond to Notch signaling. We characterized the transcriptional response to Notch activation in this cell line by performing RNA-seq on cells grown on tissue culture dishes coated with JAGGED1, a Notch ligand. Differential gene expression analysis shows induction of both canonical Notch responsive genes, as well as cell-type specific Notch responsive genes. Overall design: Cells were plated overnight on the JAGGED-coated tissue culture plates in the presence of 100 nM gamma secretase inhibitor (GSI), a small molecule that prevents the final proteolytic cleavage of the Notch receptor. The following morning, GSI was washed out by 3 media changes, and cells were collected at 2, 4, and 24 hours after GSI washout. Additionally, a mock washout control was performed in which GSI was maintained in the media and cells were harvested after the mock washout. Three replicates were collected of these 4 conditions. RNA-seq was performed on these 12 samples. Genes that were differentially expressed at 2, 4, and 24 hours were identified.