RNA was extracted from H9 hESCs; H9-derived NCSCs at D15; H9-derived NCSCs maintained for 40 additional days in E6-CSFD (D55); H9-derived pericyte-like cells at D19, D22, and D25 (three independent differentiations at the D25 time point); H9-derived pericyte-like cells maintained for 20 additional days in E6 + 10% FBS (D45); CS03n2-derived pericyte-like cells at D25; IMR90C4-derived pericyte-like cells at D25; and primary brain pericytes using the RNeasy Mini Kit (Qiagen) as described above. TruSeq stranded mRNA libraries were prepared, cDNA-synthesized, pooled, and distributed over two sequencing lanes, and samples were sequenced on an Illumina HiSeq 2500 at the University of Wisconsin–Madison Biotechnology Center. Reads were mapped to the human genome (hg38) with HISAT2 (v2.1.0), and transcript abundances (FPKM) were quantified with Cufflinks (v2.1.1). FPKM values from the two sequencing lanes for each sample were averaged. Hierarchical clustering was performed with Morpheus (https://software.broadinstitute.org/morpheus) using the one minus Pearson correlation with average linkage. GO analysis was performed using the PANTHER (76) online tool (http://pantherdb.org).

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