Extended Data Fig. 2. Transient ETV2 expression in adult human ECs is sufficient for the generation and maintenance of durable long-lasting R-VEC vessels in vitro.
a, Schematic for ETV2 mRNA and protein levels assessment at each of the three stages of R-VEC vessel formation. b, Quantification of ETV2 mRNA levels at each stage of vessel formation. c, d, Western blot analysis (c) and densitometric quantification (d) of ETV2 protein levels at each stage of vessel formation. GAPDH was used as a loading control. e, A proteasome inhibitor (MG132) restored ETV2 levels by ~sixfold when added to R-VECs during the stabilization stage. f, Densitometric quantification of western blots in e. g, qRT–PCR (g) and western blot (h) assessment of ETV2 levels after doxycycline removal. i, Representative images of GFP+ iR-VECs on Matrigel with inducible ETV2 expression at 2 months. ETV2 was turned off at day 0, day 7 and at 4-weeks post start of the remodelling stage 2. j, Quantification of iR-VEC vessels at 2 months. k, Electron microscopy pictures of a lumen present both in vessels in which doxycycline was continuously present and in vessels in which doxycycline was removed after 1 month. Data are mean ± s.e.m. NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001. For statistics, see Supplementary Data 1. For medium formulations, see Supplementary Data 2.