Fig. 3.

Wasb is required for oriented cell migration. (A) Montage of two ISV sprouts from 26 hpf to 38 hpf in MO-wasb embryo. Top panel: live imaging of Tg[fliep:nls-mCherry; fliep:lifeactGFP] MO-wasb. Black, F-actin; red, nuclei. Bottom panel: pictogram of live imaging. Red ECs represent cells in aISVs; blue ECs represent cells in vISVs. Dark red ECs indicate cell division events (time point 31:10) and emigration of cells from aISV to DLAV (time points 35:20-38:20). Dark blue and dark red cells indicate vascular remodelling (time point 31:00), cell division (time point 32:20, 33:20) and emigration of ECs towards the DLAV (time point 31:10, 33:20-38:20). Dark grey cells represent cell anastomosis (time point 28:30). (B) EC velocity (µm/h) over time (hpf). Red, velocity for ECs in aISVs; blue, velocity of ECs in vISV. Left pictogram demonstrates positive and negative velocity relative to dorsal and ventral movement, respectively. (C) Number of cells that exchange from ISVs with DLAV and fractions of ISVs that are affected (immigration from DLAV to aISV AKS test, P<0.05; emigration from vISV to DLAV AKS test, P<0.05; MO-control versus MO-wasb). (D) Number of cells that exchange from PCV with vISV and corresponding fractions of vISVs. (E) Number of mitotic events in ISVs (MO-control versus MO-wasb vISV, AKS test P<0.05). (F) Cell number in ISVs 44 hpf and distribution among ISVs (MO-control versus MO-wasb vISV, AKS test P<0.05). Grey bars (C-F) are values of MO-control. (G) Vessel diameter (µm) over time (hpf) for MO-control aISV (dotted redline), MO-wasb aISV (full redline), MO-control vISV (dotted blue line) and MO-wasb vISV (full blue line). Black vertical lines represent the equivalent time points in montage. Data are mean and 95% confidence interval.