Fig. 4

Fig. 4 VEGF regulates of SMAD3 expression levels in pericytes.

a We differentiated 2 AD and 2 control patient-derived iPSCs to pericytes as previously described⁷⁹, treated the differentiated pericytes with recombinant VEGF, VEGFR2 (KDR) inhibitor cocktail, and aggregated Aβ and analyzed the impact on SMAD3 expression at three time points (6, 12, and 24 h). b–d Validation of pericyte differentiation was performed via flow cytometry, immunocytochemistry, and RT-qPCR. We observed decreased expression of iPSC pluripotency marker, TRA-10, and increased expression of pericytic PDGFRB and NG2 through FACS in the differentiated pericytes. We also visualized and confirmed pericytic PDGFRB expression through ICC (scale bar:100 µm) and observed upregulation of pericyte and vascular markers after differentiation. Statistics: two-sided paired t-test, n = 4 biologically independent samples; within each experiment, n = 6 technical replicates. e We observed significant decrease in SMAD3 expression after 24 h of VEGF treatment. f Consistently, VEGFR2 inhibitor cocktail treatment caused significant increase in SMAD3 expression at all time points. g Aggregated Aβ treatment did not cause significant change in SMAD3 expression (n = 5 per each duration). Statistics derived from biologically independent replications (different iPSC lines and differentiation batches). All boxplots represent the first quartile, the median, and the third quartile. The upper whisker indicates the maximum value no further than 1.5 times the inter-quartile range from the third quartile. The lower whisker indicates the minimum value no further than 1.5 times the inter-quartile range from the first quartile. Source data are provided as a Source Data file. Figure 4/panel a Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs license.