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Fig. 8
fli1:etv2 DNA injection rescues arterial and venous marker expression in Vegfr inhibited embryos. (A-H) In situ hybridization analysis showing expression patterns of arterial and venous markers upon Vegfr inhibition and etv2 DNA overexpression. (A,B) Uninjected embryos treated with DMSO displayed normal expression patterns of arterial marker cldn5b (A) and venous marker flt4 (B). (C,D) fli1:etv2 DNA injection does not significantly affect endogenous vascular endothelial marker expression, but did result in small patches of ectopic expression (white arrowheads). (E,F) SU5416 inhibition results in downregulation of arterial cldn5b (E) and venous flt4 (F) marker expression. Embryos injected with fli1:etv2 DNA and treated with SU5416 have increased cldn5b (G) and venous flt4 (H) vascular endothelial marker expression, as compared to SU5416 treated embryos (E,F) demonstrating a partial rescue of Vegfr inhibition phenotype. Arrows indicate DA and black arrowheads indicate PCV. Lateral view of 24 hpf embryos, anterior to the left. Numbers of embryos showing displayed phenotype are listed. (I) qPCR analysis of arterial (cldn5b), venous (flt4, dab2, stab1l) and pan-endothelial (cdh5) marker expression. Note that etv2 overexpression under fli1 promoter partially or completely rescues all marker expression in SU5416 treated embryos. *, p<0.05; ** p<0.01; significance of expression values in fli1:etv2+DMSO embryos was compared to wt+DMSO; ±SEM is shown.
Reprinted from Developmental Biology, 424(2), Chetty, S.C., Rost, M.S., Enriquez, J.R., Schumacher, J.A., Baltrunaite, K., Rossi, A., Stainier, D.Y., Sumanas, S., Vegf Signaling Promotes Vascular Endothelial Differentiation by Modulating etv2 Expression, 147-161, Copyright (2017) with permission from Elsevier. Full text @ Dev. Biol.