Fig. 2

sec14l3/SEC14L2 are required for vascular formation in vivo and in vitro. a sec14l3-tMO injection impairs arterial-venous segregation and luminal formation in zebrafish. Embryos of Tg(kdrl:EGFP)^s843Tg;p53^+/+ (left panel) or Tg(kdrl:EGFP)^s843Tg;p53^−/− fish (right panel) were used. White and yellow brackets indicate DA and PCV respectively; red arrows indicate ISVs sprouting. The ratio in the right corner indicates the number of embryos with observed pattern/the total number of observed embryos. b sec14l3-sMO mediated knockdown of sec14l3 causes trunk vascular defects. 0.5 ng sec14l3-sMO and 150 pg sec14l3 mRNA were co-injected for rescue experiment. c Statistical analyses of DA and PCV diameters at 25 hpf in b (n = 20 embryos). d Vasculature defects in sec14l3 mutant embryos. Heterozygous mutants were intercrossed and embryos were harvested for vasculature observation and genotyping analysis individually. e Statistical analyses of DA and PCV diameters at 30 hpf in d. The total number of embryos in each genotype is indicated. f sec14l3-sMO injection causes angioblast migration defects. 0.5 ng sec14l3-sMO and 100 pg sec14l3 mRNA were co-injected for rescue experiment. WISH using etsrp probe marks the medial (arrowhead) and lateral (arrow) progenitor populations simultaneously. g Ventral sprouting defects in sec14l3 mutant embryos. h SEC14L2 knockdown inhibits the in vitro wound closure process. HUVECs infected with control or SEC14L2 shRNA were used for wound closure observation. The ratio of recovered wound width at a specific time point post-wounding to the initial wounding width was calculated. Dashed lines indicate wound edges. Statistical data are shown (n = 3). i SEC14L2 knockdown inhibits in vitro tube formation. HUVECs were infected with shRNA-packed lentivirus and the complete tube number was scored per field at 24 h after plating. The statistical data are shown on the right (n = 3 independent experiments, 60 fields total). All statistical data are shown as mean ± SEM. One-way ANOVA tests were used for statistical analyses in c, e and t-tests were used for h–i. *p < 0.05; **p < 0.01; ***p < 0.01; ns, not significant. Scale bars, 25 μm for a, 50 μm for b, d, g, and h–i, 100 μm for f. Source data are provided as a Source Data file