|
Fig. S5
Related to Figure 5. Interactions between Sema-PlxnD1 and VEGF Pathway Mutants, Effects of SU5416 on sflt1 Expression and Validation of shRNA-Mediated Knockdown of PLXND1 and FLT1 in HUVECs
(A) Cellularized SeA sprouts in WT, obd and obd; plcg1. 32 hpf trunk vasculatures. EC nuclei, green. Somite boundaries, red. Genotypes indicated on top. (B) SU5416 mediated inhibition of VEGF signaling does not visibly affect sflt1 levels in either WT or obd. 28 hpf trunks of embryos subjected to WISH with sflt1 riboprobe (blue) after treatment starting at 20 hpf with 0.025% DMSO (vehicle control) or 0.5 μM SU5416 (in 0.025% DMSO). (C) SeA phenotype of kdrl and obd; kdrl double mutants at 32 hpf. (D) Quantification of PLXND1 and FLT1 mRNA in HUVECs following treatment with control, PLXND1-targeting or FLT1-targeting shRNAs. (E) *p < .05, ***p < 0.001. Error bars: s.e.m. (C) n = 8 for kdrl, n = 8 for obd; kdrl. (A, B, C) Anterior, left; dorsal, up. Scale bars, 30 μm (A and C); 50μm in (B).
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