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. 2018 Feb 15;10(3):875–889. doi: 10.1016/j.stemcr.2018.01.009

Figure 4.

Figure 4

The Progeny of Hemangioblasts Is the Prime Target of HOXB4 Activity

(A) Ectopic HOXB4 activity was induced by addition of Tam at the indicated stages of iRunx-ESC differentiation. BL-CFC assays were performed with 10,000 FLK-1+ iRunx EBd3.5 cells each, +/− Tam.

(B) The total number of colonies was counted 4 days later, the average numbers of n = 3 independent experiments are indicated. +,+,+, Tam (HOXB4) was continuously present; −,−,−, without Tam (no HOXB4 induction); −/−/+, HOXB4 was first induced after dissociation of colonies; −/+/+, HOXB4 was first induced in EBd3.5 FLK-1-sorted cells; −/+/−, HOXB4 was only transiently induced during blast colony formation by FLK-1+ hemangioblasts.

(C) Individual colony numbers and arithmetic means (bars): −/−, no Tam; −/+, induction of HOXB4 in FLK-1-sorted cells; +/+, continuously induced HOXB4. For statistical analysis, an unpaired, two-sided t test with Welch's correction (n = 6) was performed. The p values of individual comparisons are indicated with a significance level defined as p < 0.05. To evaluate HE formation, 10,000 cells of pooled, dissociated colonies were placed onto OP9 stroma cells +/− Tam. The numbers of HE colonies grown on OP9 cells after 3 days are shown.