Fig. 7

CD146 is essential for lymphangiogenesis independent of angiogenesis. (a,b) Tg (fli1a: EGFP) embryos injected with plasmid of CD146 ∆C-GFP or its empty control, which was induced to express CD146 ∆C-GFP by 37 °C heat-shocking for 30 min. The embryos were harvested at 5 dpf. Tetramethylrhodamine-dextran was used to visualize normal TD (green arrow) and abnormal TD (green stars). The criterion used to classify the TD formation in (b) was the same as that used in Fig. 4d. Scale bars: 30 μm. (c,d) Tg (flk1: mcherry) embryos were treated as same as in (a). The embryos were harvested at 36 hpf, 48 hpf and 72 hpf, respectively. Vasculature of blood and lymphatic vessels were observed by confocal microscopy. Images were flanked by redrawing of the vessel contours. Transient lymphangiogenic structures (lymphangionenic sprouts; PL cells) were labeled light green and the other vessels (PCV, DA and ISV) were labeled dark grey. Scale bars: 20 μm. (e) Tg (fli1a: EGFP) embryos were injected with plasmid of CD146 ∆C-GFP or its control plasmid together with control or human vegfc mRNA and were heat-shocked to express CD146 ∆C-GFP at 37 °C for 30 min. Late-phase microangiography was used to visualize typical phenotype of lymphatic capillaries in embryos at 3 dpf. The typical ratio was showed in each panel. Scale bars: 50 μm.