|
Fig. 6
Notch Signaling Loss Does Not Phenocopy obd
(A and B) Expression of Notch′s activity nuclear reporter Tg(Tp1bglob:hmgb1-mCherry)jh11 (red) in the endothelium (gray) of WT (A) and obd (B).
(C–F) obd; mib. (C) Endothelium, green. SBs, red. (D–F) WISH with sflt1, flt4 and mflt1 riboprobes, as indicated. Double mutant phenotypes classed as obd-like (C and D) or mib-like (E and F) based on the mutant they resemble most. Note lack of sflt (as in Figure 3G) and ectopic aortic flt4 (yellow arrowhead; as in Figure S6A) and venous mflt1 stainings (green arrowhead, as in Figure S6B).
(G–I) Angiogenic cell abundance within the trunk′s arterial tree of WT, obd, mib (G) and obd; mib (H) in Tg(fli1:nEGFP)y7 embryos.
(G–H) EC nuclei, green. SBs, red.
(I) Quantification; n = 10 per genotype.
(J–L) SeA sprout abundance in plcg, mib; plcg, obd; plcg (J) and obd; plcg embryos injected with 10 ng of mib MO (mib MO) (K).
(J and K) Endothelium, green. SBs, red.
(L) n = 8, 7, 11 and 9 for plcg, mib; plcg, obd; plcg and obd; plcg (mib MO), respectively.
Scale bars represent 50 μm (A, B, and D–F), 30 μm (C, G, H, J, and K).
(I and L) *p < 0.05, ***p < 0.001. Error bars represent SEM.
(A–F, G, H, J, and K) Anterior, left; dorsal, up. Trunk images and quantifications: 32 hpf (A–C, G–L), 28 hpf (D–F).
Reprinted from Developmental Cell, 21(2), Zygmunt, T., Gay, C.M., Blondelle, J., Singh, M.K., Flaherty, K.M., Means, P.C., Herwig, L., Krudewig, A., Belting, H.G., Affolter, M., Epstein, J.A., and Torres-Vazquez, J., Semaphorin-PlexinD1 Signaling Limits Angiogenic Potential via the VEGF Decoy Receptor sFlt1, 301-314, Copyright (2011) with permission from Elsevier. Full text @ Dev. Cell