Molecular mechanisms underlying the differentiation of brain mural cells from neural crest are poorly understood. We found that activation of Notch3 signaling in human pluripotent stem cell-derived neural crest (using lentiviral overexpression of the human Notch3 intracellular domain, N3ICD) was sufficient to direct the differentiation of mural cells. Overall design: RNA-seq was performed on cells from the DF19-9-11T, H9, IMR90-4, and WTC11 hPSC lines. For each line, differentiation-matched samples of neural crest cells, and GFP-negative and GFP-positive cells isolated via FACS 6 days after transduction of neural crest with N3ICD-GFP lentivirus, were analyzed.