|
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 MOsb) 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 MOsb) showing pericardial edema (middle panel, arrowhead) and cranial hemorrhage (right panel, arrows).
|
