|
Fig. S2 Transgenic zebrafish reporters for endothelial microtubule and actin cytoskeletons. Related to Figure 2. (A) Tg(fli1ep:EGFP-DCX) embryo at 32hpf. EGFP-DCX allows visualization of diverse microtubule structures during cell division. A single MTOC (microtubuleorganization centre; arrow; 00:00) doubles (00:20) and relocates to opposite poles of the dividing cell (00:25 and 00:35). A microtubule-based mitotic spindle forms (arrowhead; 00:40) between the centrosomes and allocates the chromosomes to the daughter cells. The cells divide (cytokinesis) and the central spindle (white arrowhead) and the MTOCs (arrows) are visible in each daughter cell. 00:00, hours:minutes. Scale bar, 20µm. (B and C) Tg(fli1ep:EGFP-Actin) embryos at 3dpf. EGFP-Actin highlights F-actin cables at cell junctions (arrowheads). ISV, intersegmental vessel; DLAV, dorsal longitudinal anastomotic vessel; ISLV, intersegmental lymphatic vessel; DA, dorsal aorta; DV, dorsal vein; VV, ventral vein. Scale bars, 10µm. (D and E) Fluorescence recovery curves of EGFP-Actin at junctions after DMSO or SMIFH2 treatment. (F) Mobile fraction and half-life of EGFP-Actin at DA or PCV junctions after DMSO or 10µM SMIFH2 treatment. Data represent mean ± SD.
Reprinted from Developmental Cell, 32, Phng, L.K., Gebala, V., Bentley, K., Philippides, A., Wacker, A., Mathivet, T., Sauteur, L., Stanchi, F., Belting, H.G., Affolter, M., Gerhardt, H., Formin-mediated actin polymerization at endothelial junctions is required for vessel lumen formation and stabilization, 123-32, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell