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Fig. S1 The angioblasts forming the bilateral cardinal veins and the common cardinal veins (CCVs) are specified as a second population and at a separate time point to the population forming the lateral dorsal aortae. (A-J) Confocal projections of Tg(fli1ep:GAL4FF)ubs4 (UAS:KAEDE)rk8 embryos at the indicated time points. Endothelial cells (ECs) (and blood cells) were visualized by transgenic Kaede expression. (A-G) Dorsal views; (H-J) lateral views. (A) EC-specific Kaede expression before photoconversion at 17 hpf, before the second population of angioblasts starts to express Kaede. (B) EC-specific non-converted Kaede (green) expression after photoconversion, indicating that not all Kaede protein was converted. (C) EC-specific converted Kaede (red) expression after photoconversion. (D) Merged image of converted and non-converted Kaede expression after photoconversion. (E) Non-converted Kaede expression, reflecting newly synthesized Kaede protein, imaged 9 hours after photoconversion. Green Kaede protein can be detected in the LDA and the bilateral cardinal veins as well as in the CCVs. (F) Nine hours after photoconversion, converted Kaede (red) expression can only be detected in the LDA, but not in the veins. (G) Merged image of the red and green channel showing Kaede expression 9 hours after photoconversion. Note that the transgenic Kaede expression can also be detected in blood cells. (H) Non-converted Kaede expression 9 hours after photoconversion is detected in the LDA and the bilateral cardinal veins as well as in the CCV. (I) Converted Kaede expression 9 hours after photoconversion is only detected in the LDA. (J) Merged image of the red and green channel showing Kaede expression 9 hours after photoconversion. Note that the transgenic Kaede expression can also be detected in blood cells.