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Fig. 3

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ZDB-IMAGE-170413-55

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Figures for Carroll et al., 2014

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Figure Caption

Fig. 3

E2 Exposure Impairs Formation of the Hemogenic Endothelial Niche

(A) E2 had a biphasic effect on HSPCs: treatment during hemogenic niche specification (12–23 hpf) decreased runx1, whereas exposure during HSC specification and budding (26–36 hpf) increased HSPCs (n ≥ 75/treatment).

(B) Treatment with E2 had no impact on axial vessel formation as assessed by tie2:GFP (n ≥ 20).

(D) Arterial ephrinb2 was robustly decreased by E2 (77/80); red arrowheads indicate artery, and blue arrowheads indicate vein.

(E) Venous flt4 was increased after E2 exposure (40/53).

(F) scl:GFP was reduced by E2 treatment (25/30).

(G) FACS of flk1:GFP+ embryos (n = 32) revealed no change in endothelial cell number by E2 (error bars indicate SD).

(H) qPCR confirmed decreased expression of flk1, ephrinB2, and scl following E2 treatment (mean of triplicate experiments ± SEM; one-tailed t test; flt4^∗p < 0.05, flk1^∗∗p < 0.01, and ephrinB2^∗∗∗p < 0.001).

(I) Injection of scl mRNA increased runx1 expression in the AGM and rescued loss following E2 treatment (n > 55 per treatment).

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Acknowledgments

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Reprinted from Developmental Cell, 29, Carroll, K.J., Esain, V., Garnaas, M.K., Cortes, M., Dovey, M.C., Nissim, S., Frechette, G.M., Liu, S.Y., Kwan, W., Cutting, C.C., Harris, J.M., Gorelick, D.A., Halpern, M.E., Lawson, N.D., Goessling, W., North, T.E., Estrogen defines the dorsal-ventral limit of VEGF regulation to specify the location of the hemogenic endothelial niche, 437-53, Copyright (2014) with permission from Elsevier. Full text @ Dev. Cell