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. Author manuscript; available in PMC: 2017 Aug 30.
Published in final edited form as: Cell Rep. 2017 Aug 15;20(7):1597–1608. doi: 10.1016/j.celrep.2017.07.067

Figure 6. HOPX is functionally required for primitive hematopoiesis.

Figure 6

(a) Fate specification of CD31 endothelial cells is not effected by HOPX KO. (b) HOPX KO does not impact functional endothelial lumen formation assays in collagen. Scale bars = 200 μm. (c–e) Hematopoiesis analysis of CD43+/CD235a+ derivatives in day 5 H-ECs with representative FACS plots (c) and colony forming assays in methylcellulose assaying for Ery-P (d) and macrophage CFUs (e) in WT vs. HOPX KO cells. Representative CFU images are shown to the right for primitive erythroid (Ery-P), and macrophage (Mac) colonies. (f–g) Quantitative RT-PCR analysis of HOPX and genes involved in hematoendothelial cell fate specification (SCL) and function (GATA1 and RUNX1) (f) as well as Wnt/β-catenin target genes AXIN2 and TROY1 in WT vs. HOPX KO cells (g). (h) Wnt-dependent regulation of CD43+/CD235a+ primitive hematopoiesis assessed by treatment of cells with the Wnt agonist CHIR-99021 (1 μM) or the Wnt antagonist XAV-939 (1 μM). (i) Differentiation of HOPX KO and WT cells into H-ECs showing that addition of the Wnt inhibitor XAV-939 (1 μM) partially rescues hematopoietic deficiencies observed in HOPX KO cells. n=4–8 per group. Values are presented as mean ± sem. *P<0.05 vs. WT. # P<0.05 vs. HOPX KO cells.