Quantification of the anti-angiogenic effects in zebrafish embryos. Stalks length (A) and number (B) were measured for fli1a:EGFP zebrafish embryos at 75hpf treated with vehicle, Thalidomide or compounds at different concentrations. The box and whiskers plots show the data points quantified for at least ten embryos per condition. (C) Representative confocal images of zebrafish for each condition. White arrows show a missing stalk, a reduction in vessel length or loss of vascular connectivity. (D) The length of stalks was plotted here for three different groups of stalks. Statistics were calculated by one-way ANOVA. n.s. = not significant; *p < 0.05; ^∗∗p < 0.01; ***p < 0.001; ****p < 0.0001.

Quantification of the teratogenic side effects in zebrafish embryos. (A) Malformations were measured by three teratogenic parameters in the zebrafish embryo: the eye area (A), the pectoral fin length (B) and the ratio between the long axis of the otic vesicle and the body length (C). The eye area (A) and the pectoral fin (B) were measured at 75hpf and the otic vesicle (C) was measured at 30hpf for optimal visualization. Lateral view of embryo eye (A), dorsal views of pectoral fins (red arrowheads in B) and lateral view of the otic vesicle for each condition are showed. For each parameter, the data are shown as percentage variation in respect to the treatment with vehicle. The data are plot in a scattered dot plot showing the mean ± SD. Statistics were calculated by one-way ANOVA. n.s. = not significant; *p < 0.05; **p < 0.01; ****p < 0.0001.