Figure 3.

Phenotypic characterization of iSMCs and pSMCs in hemodynamic cocultures. iECs and pECs were cocultured with iSMCs and pSMCs, respectively, and exposed to CCA hemodynamics or static conditions. (A): Scatterplot representation of differentially expressed genes (DEGs) in pSMCs and iSMCs. Each point represents log2fold‐change of a DEG for each cell type in hemodynamic coculture versus cell‐type‐matched static control coculture. A purple DEG exhibits the same directionality in fold‐change for both cell types, while a green DEG exhibits opposite directionality; color intensity increases with statistical significance. The response similarity index (RSI) value is a single numerical value that incorporates the fold‐change and significance for each DEG in both cell types. A pathway‐weighting approach also identified several biological themes that emerged from the RSI analysis. (B): Gene expression for pSMCs and iSMCs of contractile SMC markers are represented as log2fold‐change of CCA hemodynamics versus static (red = upregulation, blue = downregulation). (C): Transcriptomic data for pSMCs and iSMCs exposed to CCA hemodynamics of contractile SMC markers are also represented as total CPM for each gene. (D): Representative photomicrographs of iSMCs and pSMCs exposed to CCA hemodynamics and stained with F‐actin (green—left panels), smooth muscle alpha actin (SMAA; red—left panels), calponin (green – mid panels), transgelin (SM22; red—left panels), and TO‐PRO‐3 for nuclei (blue—all panels). Scale bar = 100 μm. Abbreviations: CCA, common carotid artery; CPM, counts per million; iSMCs, iPSC‐derived SMCs; pSMCs, primary SMCs; SMAA, smooth muscle alpha actin.