Fig. S3

Knockdown of Rasa3 in Zebrafish induces tubulogenesis defects in the trunk vasculature.

A. Detection of Rasa3 level in lysates from Control (Ctl) and Rasa morphant embryos. GAPDH was used as control. B. General morphology of Ctl and Rasa3 morphant embryos. C. Tg(fli1a:eGFP)y1 embryos were injected with control morpholino (MoCtl) or with morpholino targeting Rasa3 (MoRasa3). Confocal pictures of the trunk vasculature were taken at 48 hpf. Ctl embryos present normal ISVs and DLAVs with open lumen (arrow head) whereas Rasa3 morphant embryos show thinner, non-lumenized vessels (arrows). Bars = 50 mm. ISV, intersegmental vessel; DLAV, dorsal longitudinal anastomotic vessels. D. Quantification of lumenized ISVs in Ctl and Rasa3 morphant embryos at 48 and 72 hpf. The p values are shown (Fisher's exact test). Results are mean from 10 ISVs/embryo in 50 embryos at 48hpf and 25 embryos at 72hpf. E. Heart rates in 32 Ctl and 30 Rasa3 morphant embryos. Histograms are mean ± SD from 35 embryos. The p value is shown (Fisher's exact test). F. Rescue experiment using GGTI298 (10 μM). Quantification of lumenized ISVs in Ctl and Rasa3 morphant embryos at 48hpf. The p values are shown. Results are mean from 10 ISVs/embryo in 35 embryos.