Fig. 5

Fig. 5 reg6 mutants have defects during anastomosis and plexus formation. Time-lapse images of wild-type TG(fli1:EGFP)^y1 (A–D) and reg6;TG(fli1:EGFP)^y1 (E–H) regenerates show that reg6 regenerates fail to complete anastomosis before outgrowth and later fail to form plexuses during blood vessel regeneration. (A) and (E) show 19-hpa regenerates (33°C) at which stage the wild-type regenerates (A) have finished anastomosis (red arrows), while reg6 regenerates failed to do so (E, yellow arrows). By 44 hpa (33°C), the regenerating vessels form plexuses in wild-type regenerates (B), but those in reg6 mutants grow out without forming plexuses. Instead, they form blisters with few branches (F). In 63-hpa reg6 regenerates (33°C), some blisters that developed at the distal ends of regenerating vessels (G) are filled with blood cells (red arrow in H) and some remain clear (black arrow in H), which can easily be perceived in bright field. Similar blisters are typically never seen in wild-type regenerates at the same stages (C, D). Insets in (C) and (G) are higher magnification images to show distance between nuclei (green foci) of endothelial cells in wild-type and reg6 regenerates, respectively. Red arrowheads point to amputation planes. Scale bar, 100 μm for (A, E); 200 μm for (B, F); 300 μm for (C, D, G, H).