Figure 4.

HEB^−/− Mesodermal Precursors Are Not Impaired in Their Ability to Give Rise to CD34⁺ Endothelial Cells

(A) Experimental overview for KDR⁺ cell isolation and reaggregation.

(B) Percentage of KDR⁺ cells in day-4 (d4) EBs derived from WT, KO-4, or KO-8 hESCs, before sorting.

(C) Pre-sort and post-sort purity of KDR⁺ cell enrichment after each sorting step (magnetic-activated cell sorting [MACS] followed by fluorescence-activated cell sorting), as determined by flow cytometry.

(D) Percentages of CD34⁺ cells in cultures at 2-day intervals after reaggregation of d4 EB-derived KDR⁺ cells. Representative flow-cytometry plots of CD34 and KDR expression are shown on the right.

(E) Experimental overview for endothelial tube formation in vitro.

(F) Images of endothelial vessel-like structures. Scale bar, 100 μm.

Error bars represent mean ± SD (n = 3 independent experiments). ^∗∗p < 0.01, ^∗∗∗p < 0.005 by Student's t test. Plot in (B) is compiled from six independent experiment, with one replicate per experiment, and plots in (C) and (D) and images in (F) are representative of three independent experiments.