ZET results for TTZ. (A) Dose-response curves showing survival and occurrence of any malformation induced by 96 h TTZ exposure, X-axis is logarithmically scaled. (B) A control embryo at 4 dpf showing normal morphology. (C) Hemorrhagic sites (arrowheads) induced by TTZ exposure. (D) Percentages of surviving embryos with different phenotypes at 4 dpf (Standard Deviations were excluded for visual clarity). (E) Severe deformities at high TTZ concentration.

Vascularization analysis on transgenic fluorescent zebrafish lines. (A) Cephalic vessel and caudal venous plexus vascularization are dose-dependently affected by TTZ; Vessels appear to be increasingly merged/clumped together instead of forming distinguishable fine structures; Notably , the three-longitudinal-vessel pattern in the CVP gradually merged (5 g/L) into a two-vessel pattern (10 g/L). (B) Normal (basket-shaped) and ectopically sprouted (arrowheads) SIV formation. (C) Percentages of ectopic sprouting at different TTZ concentrations.

TTZ inhibits HUVEC angiogenic activities in a dose-dependent manner. (A) TTZ inhibits HUVEC proliferation in the BrdU assay. (B-D) TTZ treatment leads to reduced tube extension and branching during tubulogenesis. (E-F) Wound healing assay showing anti-migratory effect of TTZ. Concentrations are log-scaled , and biological replicates are represented using different colors. Asterisks denote statistical significance: *p < 0.01; **p < 0.001.