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Fig. 4
GB9 treatment inhibits the growth of ISV cells. (A–D) At 30 hpf, TUNEL labeling (in wild-type AB strain) and AO staining (in Tg(kdrl:mCherry) embryos) were used to detect apoptotic cells (arrows in (A,B) and green spots in (C,C’,D,D’)) in 0.3% DMSO control (A,C,C’), and GB9-treated embryo (B,D,D’). Some increased apoptotic cells were observed on the skin close to tail region in GB9 treated embryo, but not in vascular regions compared to untreated controls (yellow arrows in (C,C’,D,D’)). At 30 hpf, ISVs show slower growth at mid-somite in the GB9 treated Tg(kdrl:mCherry) embryos (hollow arrowheads in (D)) and less endothelial cell sprouting and loop formation at CVP (hollow arrow in D) compared to control embryos (arrowheads and arrow in (C). The number of cells forming each ISV counted in control Tg(kdrl:mCherry; fli1a:negfp y7) (E) and GB9-treated embryos (F) at 30 hpf. (G) Quantification of average endothelial cells per ISV is 3.9 ± 0.3 (ISV n = 60) and 2.2 ± 0.5 (ISV n = 80) in control and GB9 treatment respectively. ** refers to p < 0.001 by an unpaired Student’s t-test. Scale bars are 100 μm for (A–F).