Fig. 8: Organ-specific transcription factors reprogram endothelial cell gene expression and alter barrier functionality.
a, Experimental workflow for HUVEC transduction with organ-specific transcription factor cocktails and subsequent functional analyses. b, Quantification of TEER across an endothelial monolayer over time, with T0 denoting the day of lentiviral-mediated transduction of the indicated transcription factors. c, Quantification of FITC-conjugated 40 kDa dextran leak across an endothelial monolayer in a Transwell permeability assay following lentiviral transduction with the indicated transcription factors. d–f, qRT–PCR analysis of the induction of BBB-related endothelial markers (d), genes encoding junctional proteins (e) and sinusoidal liver endothelial transcripts (f) following transduction of HUVECs with the indicated transcription factors. Expression is relative to non-transduced cells. Non-trans, non-transduced. g, Representative images of HUVECs immunostained for the junctional proteins VE-cadherin (cyan) and Claudin5 (yellow), with nuclei labeled by DAPI (gray) staining, and visualization of the mCherry reporter (magenta) following transduction with the indicated transcription factor(s). There were three biological replicates, each containing three technical replicates, for each experiment. Data shown are mean ± s.d. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (two-way ANOVA with Tukey’s multiple-comparison test). Panel a created with BioRender.com.
