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Fig. 1. Characterization of methylcellulose colonies. (A) Micrographs of
representative colonies from methylcellulose cultures: 2° EB colony (A
'), transitional colony (Trans-CFC; D') and hemangioblast colony
(BL-CFC; G'). Microwells of colonies replated on matrigel: 2° EB
with adherent mesodermal cell type (B',C'); two independent
transitional colonies showing mostly hematopoietic cells (E',F');
BL-CFCs showing both adherent endothelial cells (arrows) and overlaying
non-adherent hematopoietic cells (arrowhead; H',I'). (B) Single
hemangioblasts were replated on matrigel for one week and treated with
fluorescent DiI-Ac-LDL. Phase contrast (left panel) and fluorescent micrograph
(right panel) images are shown. (C) RNA from individual
Arnt+/+ and Arnt–/– 2°
EB, Trans-CFC and BL-CFC colonies were analyzed for the expression of the
indicated genes by radioactive RT-PCR: ß-actin (control), Rex1
(ES cells), Bry (mesoderm), Scl (endothelial/hematopoietic
cells) and GATA1 (hematopoietic cells). As BL-CFCs are generally smaller
colonies, the signal for ß-actin is lower.
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