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Fig. 6
Angiogenic phenotype rescue with rat-BG mRNA. (a) Images correspond to lateral views of representative Tg(fli:EGFP)y1 embryos injected with 7 ng mo-BG02-mis (CONTROL), 7 ng mo-BG02 (mo-BG02), or 7 ng mo-BG02 together with 250 pg of in vitro transcribed rat-BG mRNA (mo-BG02 + rat-BG mRNA). Asterisks indicate examples of incomplete ISV. (b) Quantitative analysis of mo-BG02 effects and rat-BG mRNA rescue of ISV development. The number of ISVs that reached the dorsal aspect of the embryo (“complete ISV”) was counted in each embryo at 48 hpf, and the result was plotted as the proportion of all, complete and incomplete, ISVs. This ratio was close to one in control embryos (0.978 ± 0.005, N = 17), and in it was significantly reduced mild-morphants injected with 7 ng of mo-BG02 (0.393 ± 0.020 N = 28, two experiments, P < 0.0001). In embryos injected 250 pg of the rat-BG mRNA together with mo-BG02, the proportion of complete ISV significantly increased (0.6191 ± 0.01852 N = 35, two experiments, P < 0,001). (c) Quantitative analysis of mo-BG02 effects and rat-BG mRNA rescue of CVP development. The mean area (µm2) occupied by ECs in the ventro-caudal region of the Tg(fi1-eGFP)y1 line was measured at 2 dpf taking the YSE as the anterior limit and caudal aorta as dorsal limit, and plotted as Area CVP. In control embryos this area was (85610 ± 2329 N = 17), while the in mild morphant embryos injected with 7 ng mo-BG02, it was significantly reduced (46250 ± 1983 N = 27, two experiments, P < 0,001). This effect was partially rescued by rat-BG mRNA, resulting in the increased area of CVP compared to morpholino alone (70350 ± 1626 N = 37, two experiments, P < 0,001).