Fig. 2

Expression of hematopoietic markers are reduced by eaf1 depletion. (A) significant reductions of erythrocytic markers hbae1 (a and b), hbae3 (c and d) and hbbe1 (e and f) were detected in eaf1-knocked down embryos at 48 hpf; expression of alas2 was also decreased in the ICM of eaf1-knock down embryos at 30 hpf (g and h). (B) The myelomonocytic marker l-plastin was reduced in eaf1-knock down embryos at 22 hpf and 48 hpf. (C) The granulocyte-specific marker mpo was reduced in eaf1-knocked down embryos at 24 hpf and 48 hpf. (D) Expression of definitive hematopoiesis marker runx1 was sharply diminished in the ventral wall of dorsal aorta in eaf1 morphants at 31 hpf, 36 hpf and 48 hpf (a–h); semi-quantitative RT-PCR assays confirmed that expression of runx1 (i) was reduced in eaf1 morphants, but was recovered by the co-injection of eaf1 mismatch mRNA. (E) Expression of definitive hematopoiesis marker c-myb was sharply diminished in the ventral wall of dorsal aorta in eaf1 morphants at 31 hpf, 36 hpf and 48 hpf (a–h); semi-quantitative RT-PCR assays confirmed that expression of c-myb (i) was reduced in eaf1 morphants, but was recovered by the injection of eaf1 mismatch mRNA. (F) The lymphoid marker rag1 was abolished in eaf1 morphants at 3.5 dpf (a and b) and 4.5 dpf (c and d). (G) The co-injection of eaf1 mismatch mRNA rescued rag1 expression at 4.5 dpf (a–c). (H) The lymphoid marker rag2 was abolished in eaf1 morphants at 3.5 dpf (a and b) and 4.5 dpf (c and d). (I) The co-injection of eaf1 mismatch mRNA rescued rag2 expression at 4.5 dpf (a–c). Eaf1–MO3, 4 ng/embryo.