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. 2015 Aug 1;29(15):1618–1630. doi: 10.1101/gad.263210.115

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© 2015 Koltowska et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 5. — mafba-dependent endothelial genes are Vegfc-dependent, and mafba is up-regulated by Vegfc signaling. (A) Schematic summary of RNA-seq from FACS-sorted sibling and mafba^uq4bh mutants at 48 hpf. n = 23 down-regulated genes; n = 68 up-regulated genes. P < 0.01 using EdgeR analysis. (B) qPCR validation of RNA-seq. RNA-seq relative changes and qPCR fold changes are displayed side by side for a subset of up-regulated and down-regulated genes. Mean ± SEM. The boxed area contains down-regulated genes. (C) Fifteen down-regulated genes analyzed by qPCR in FACS-sorted ECs from 48-hpf vegfc-induced and 30-hpf MO-vegfc embryos. Reductions in expression were observed in both mafba^uq4bh and MO-vegfc embryonic endothelium. Mean ± SEM. (D) Expression of mafba in control uninjected (n = 30/33), MO-vegfc-injected (n = 18/18), and MO-vegfr3-injected (n = 33/33) embryos at 48 hpf. Enriched dorsal PCV expression (arrows) is absent in MO-vegfc and MO-vegfr3 embryos (brackets). Bars, 100 μm. (E) qPCR validation of mafba reduction in FACS-sorted ECs from MO-Vegfc embryos. Expression is given relative to the geometric average of kdrl, cdh5, and lyve1b expression. Mean ± SEM. (F) Expression of mafba in control and vegfc-induced [Tg(prox1a:Kalt4);Tg(10xUAS:Vegfc)] embryos. mafba expression is induced throughout the PCV in vegfc-induced embryos. n = 16/16. ISH staining was for shorter periods than in E to highlight increased expression. Bars, 100 μm. (G) qPCR validation of mafba induction in FACS-sorted ECs from Tg(prox1a:Kalt4);Tg(10xUAS:Vegfc) embryos. Expression is given relative to the geometric average of kdrl, cdh5, and lyve1b expression. Mean ± SEM.