|
|
Blood, Vol. 108, Issue 1, 160-166, July 1, 2006
Zebrafish G protein  2 is required for VEGF signaling during angiogenesis
Blood Leung et al.
108: 160
Supplemental materials for: Leung et al
Files in this Data Supplement:
- Figure S1. Translational blocking by gng2-ATG morpholino detected by in vitro cell lysate assay (JPG, 81.9 KB)
-
(A) Translational blocking morpholino, targeted against the 5'UTR spanning the ATG start codon of gng2 open reading frame, fused to V5-6XHistidine epitope. (B) Dose dependent inhibition of gng2 translation by gng2-ATG morpholino using in vitro cell lysate. No inhibition by 5-base mismatch control morpholino. Anti-V5 antibody was used to detect the in vitro translation of gng2-V5 fusion protein. (C) WT control zebrafish embryo at 1 dpf. (D) gng2-ATG-MO knockdown embryo at 1 dpf. All scale bars are 100 µm (C-D).
- Figure S2. List of supplementary videos 1-4 (JPG, 113 KB)
-
Loss-of-function of gng2 led to blood flow defect at the intersomitic vessels in developing zebrafish embryos. Blood circulation in the vasculatures was monitored as a functional assay of vessel development in zebrafish embryos. Various morpholinos against gng2 were microinjected into 1-cell stage zebrafish embryos. The first morphological sign of angiogenic defect was detected at about 3 dpf when the knockdown embryos showed reduced or missing blood flow to the intersomitic vessels, although there was blood circulation in the dorsal aorta and cardinal vein. The intersomitic vessels are particularly interesting since they develop by sprouting from the dorsal aorta and cardinal vein, a process that resembles angiogenesis in physiological and pathophysiological processes. Videos 1-4 are enlarged trunk regions. All scale bars are 100 µm.
- Figure S3. Whole-mount in situ analysis of gng2 expression from 18hpf to 108 hpf in zebrafish embryos (JPG, 94 KB)
-
gng2 expression in the central nervous system (CNS) and axial vascular tissues at 18hpf to 24hpf zebrafish embryos, its expression gradually reduced in the axial vascular tissues but remained in the CNS from 28hpf. (v.m.h. ventral mid- and hind-brain; tel. telecephalon; a.v. axial vasculature; s.n. spinal cord neurons). All scale bars are 100 µm.
- Video 1. Blood circulation in WT control zebrafish embryo at 3 dpf (AVI, 5.86 MB)
-
Notice the blood flow in the intersomitic vessels.
- Video 2. Blood circulation in gng2-MO splicing knockdown embryo at 3dpf (AVI, 5.84 MB)
-
Showing blood flow in the dorsal aorta and cardinal vein but the flow to the intersomitic vessels was blocked or reduced.
- Video 3. Blood circulation of gng2-ATG-MO translational blocking morpholino knockdown embryo at 3dpf (AVI, 5.73 MB)
-
Showing blood flow in the dorsal aorta and cardinal vein but the flow to the intersomitic vessels is blocked or reduced.
- Video 4. Blood circulation in 5-base mismatch control morpholino (5-mismatch-MO) knockdown embryo at 3dpf (AVI, 5.76 MB)
-
Notice the normal blood flow in the intersomitic vessels.
|
|
