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Fig. 1. Characterization of the embryonic blood phenotype in cia. (A-L) Ventral views of the anterior region of embryos. (A-C,G-I) Whole-mount o-dianisidine staining of wild-type and cia embryos. Compared with wild type at 36 hpf (A), cia^hp327 (B) (shown as representative of cia^hs019 and cia^iu089 at all stages) lack hemoglobinized erythrocytes, while cia^tu25f (C) manifest a moderate decrease. At 72 hpf, circulating hemoglobinized erythrocytes are still absent in cia^hp327 (H) and a moderate decrease is again observed in cia^tu25f (I) compared with wild type (G). (D-F,J-L) Whole-mount RNA in-situ hybridization for ße1 globin in wild-type and cia embryos. (D-F) At 36 hpf, cia embryos are indistinguishable from wild type, while the onset of anemia in cia is apparent at 72 hpf, with cia^hp327 (K) possessing less than approximately 30% of cells compared with wild type (J), and cia^tu25f (L) exhibiting an approximate 50% decrease in erythrocytes.

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