Fig. S4

The Tg(efnb2a:eGFP) transgenic reporter is not responsive to nr2f2. (A) Transmitted light image of a 24 hpf Tg(efnb2a:eGFP) transgenic animal. (B) Green fluorescence confocal image of GFP expression in the animal in panel A, showing expression in muscle, neural, vascular, and other tissues. The red box shows approximate area magnified in panel C. (C) Higher magnification green fluorescence confocal image of GFP expression in the dorsal aorta (DA) but not in the posterior cardinal vein (PCV) in the animal in panel A. (D) Schematic diagram of a 1 dpf zebrafish (modified from (Kimmel et al., 1995)) with a red box showing the approximate position of images in BF. (E-I) Green fluorescent confocal micrographs of 24 hpf Tg(efnb2a:eGFP) transgenic zebrafish. Image planes above and below the dorsal aorta and intersegmental vessels were not used in the reconstructed confocal images shown here, to eliminate obscuring somitic expression of the transgene and permit visiualization of the vessels. Animals in image panels E-I were either untreated (E), injected with nr2f2 MO (F), injected with kdrl-CF (Full length nr2f2 under the control of the kdrl promoter; G), injected with kdrl- ECt (chimeric protein with nr2f2 lacking the transactivation domain fused to engrailed repressor domain; H), or injected with kdrl-VCt (chimeric protein with nr2f2 lacking the transactivation domain fused to VP16; I). Scale bar, 100 µm.