Fig. 3

Vegfaa and Vegfab are necessary for islet vessel development. (a) RT-PCR of vegfaa and vegfab on sorted 2 dpf, 3 dpf, and adult beta-cells. Full length gel is presented in Supplementary Fig. 3. (b) Phenotypic score of islet vessels in non-injected, control morpholino and vegfaa, vegfab, or vegfaa/vegfab morpholino injected Tg(fli1:EGFP; insa:tagRFP) embryos. Phenotypes are scored such that no phenotype is comparable to wildtype (more than 7 endothelial cells adjacent to beta-cells), mild phenotype (4–7 endothelial cells adjacent to beta-cells), and severe phenotype (less than 4 endothelial cells adjacent to beta-cells). (c) The number of beta-cells in 72 hpf Tg(fli1:EGFP; insa:tagRFP) control and morpholino injected embryos. In the vegfaa, vegfab, and vegfaa/ab morpholino injected embryos, only the embryos that demonstrated a reduction or absence of islet vessels were counted. n = 9–21. Box-and-whisker plots show median, and circles represent individual zebrafish. (d–h) Confocal projections of (d) non-injected, (e) scrambled injected, (f) vegfaa, (g) vegfab, or (h) vegfaa/ab morpholino injected embryos at 72 hpf; endothelial cells (green), beta-cells (red), and DAPI (grey). Scale bar = 10 μm.