ZFIN Image: Fish et al., 2008, Fig. S9

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Figure Caption

Fig. S9 Spred1 and VEGF Signaling Are Required for Vascular Integrity and Function
(A) Treatment of 48 hpf embryos with 5 μM of a VEGF receptor inhibitor, Vatalanib, for 18 h results in a reduction in lumen diameter in branchial arches (arrows).
(B) spred1 expression was quantified in FACS-isolated GFP⁺ endothelial cells from 72 hpf Tg(flk1:GFP)^s843 zebrafish embryos by real-time PCR. Shown is the amplification curve for spred1 and the endogenous control gene tbp.
(C) spred1 expression was inhibited by injection of a MO (Spred1 MO^sb) that was designed to cause exon 2 skipping and the production of a premature stop codon and nonsense-mediated decay of spred1. Shown is an agarose gel of RT-PCR using primers in exon 1 and 3 (left) and qRT-PCR (right) using primers in exon 4 and 5 in 72 hpf embryos.
(D) Lateral view of 72 hpf zebrafish embryos injected with splice-blocking Spred1 MO (Spred1 MO^sb) showing pericardial edema (middle panel, arrowhead) and cranial hemorrhage (right panel, arrows).

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Acknowledgments

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Reprinted from Developmental Cell, 15(2), Fish, J.E., Santoro, M.M., Morton, S.U., Yu, S., Yeh, R.F., Wythe, J.D., Ivey, K.N., Bruneau, B.G., Stainier, D.Y., and Srivastava, D., miR-126 regulates angiogenic signaling and vascular integrity, 272-284, Copyright (2008) with permission from Elsevier. Full text @ Dev. Cell