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Fig. 1 bcas2 is expressed in the intermediate cell mass (ICM) and required for primitive hematopoiesis. (A) Whole-mount in situ hybridization (WISH) assay showing bcas2 expression in the ICM at the 18-somite stage and 22 hpf. The dotted lines represent the section position and the black arrowheads indicate the ICM region. n, notochord. (B) Double fluorescence in situ hybridization (FISH) assay showing the expression pattern of bcas2 and gata1 in the ICM at 22 hpf. (C, D) Comparison of bcas2 expression in cloche mutants (C) or bcas2 heterozygous mutants (D) along with their corresponding siblings. (E, F) Expression analysis of gata1 and hbbe3 in bcas2+/Δ7 and bcas2+/Δ14 embryos. (G) Hemoglobin detection using o-dianisidine staining in bcas2 homozygous mutant at 36 and 48 hpf. (H) Representative images of yolk sac from the hemangioblast-specific Bcas2 knockout mice and their siblings. Bcas2F/F females were crossed with Bcas2F/+;Kdr-Cre males to induce the deletion of Bcas2 in hemangioblasts. Scale bars, 100 μm (A, C–G), 50 μm (B), 1 mm (H).