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ZFIN ID: ZDB-FIG-101123-16
Murphy et al., 2010 - Disruption of angiogenesis and tumor growth with an orally active drug that stabilizes the inactive state of PDGFR{beta}/B-RAF. Proceedings of the National Academy of Sciences of the United States of America   107(9):4299-4304 Full text @ Proc. Natl. Acad. Sci. USA
Anatomical Term:
Stage: Long-pec

Fig. 4 Combination of imatinib and GW 5074 inhibits angiogenesis similarly to compound 6. (A) Effect of 3, 6, vatalanib, imatinib, GW 5074, and a combination of imatinib (PDGFR inhibitor) and GW 5074 (RAF inhibitor) on intersegmental vessel formation in Tg:(fli1-egfp) zebrafish embryos. Embryos were treated with DMSO, 5 μM 3, 5 μM 6, 1 μM vatalanib, 5 μM imatinib (Im), 1 μM GW 5074 (GW), and the combination of 5 μM imatinib and 1 μM GW 5074 (Im + GW) from 16 until 48 hpf. Z stacks from laser-scanning confocal microscopy are shown depicting formation of the intersegmental vessels at 48 hpf. n = 6 embryos/treatment. (Scale bar, 100 μm.) (B) HUVECs were cocultured with hTERT-human hepatic stellate cells in a 3D collagen matrix in the presence of complete EBM-2 medium to monitor pericyte-associated endothelial tube formation. The stellate cells were labeled with 10 μg/mL red fluorescent dye [DiIC(3); BD Biosciences] for 1 h before the start of the experiment. Inhibitors were added to the cocultures 6 h postseeding at the following concentrations: DMSO, 2.5 μM 3, 2.5 μM 6, 1 μM imatinib (Im), 0.5 μM GW 5074 (GW), and the combination of 1 μM imatinib and 0.5 μM GW 5074 (Im + GW). The endothelial tubes were stained at 24 h by adding 2 μL FITC-labeled Ulex europaeus lectin (Vector Labs) per well. Images were acquired 48 h postseeding of the cells. One representative panel from three independent experiments is shown. Green, FITC-lectin-labeled endothelial cells; red, DiIC(3)-labeled stellate cells. Inset in each panel displays a higher-magnification view of the endothelial cell/stellate cell interactions. (Scale bar, 200 μm.) (C) Tube lengths were measured using MetaMorph software for each tube for all 10 fields that were acquired. The % pericyte-covered tube length was calculated from the ratio of tube length sums for the tubes with and without pericyte contact. Error bars are reported as ± SEM of two wells per group. *P < 0.05 compared to DMSO group.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP y1Tg control Long-pec vasculature IFL
y1Tg chemical treatment: pharmaceutical Long-pec vasculature IFL
Antibody Labeling Details No data available
Phenotype Details No data available
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA