Knockdown of aggf1 expression results in abnormal primitive hematopoiesis during zebrafish embryogenesis. (A-C) Dramatic reduction of the number of erythroid cells by an aggf1 morpholino (MO1). Zebrafish embryos in the 1- to 2-cell stage were injected with 2 ng of Std-MO (A) or 2 ng of MO1 (B). Erythroid cells in the circulatory system were detected with o-dianisidine at 48 hpf. A similar experiment with injection of 2 ng of MO1 together with 100 pg of zebrafish aggf1 mRNA (C) into embryos was performed to determine whether overexpression of aggf1 can rescue the defects in aggf1 morphants. MO1 was designed in a manner to knock down endogenous zebrafish aggf1 mRNA only and does not knock down the in vitro synthesized aggf1 mRNA used for injection. (D-F) Knockdown of aggf1 reduces the gata1 signal, a marker for erythroid progenitors. Zebrafish embryos (1- to 2-cell stage) were injected with 2 ng of Std-MO (D), 2 ng of aggf1 MO1 (E), or 2 ng of aggf1 MO1 together with 100 pg of aggf1 mRNA (F) and used for WISH at 26 hpf with an antisense probe for gata1. (G-I) Knockdown of aggf1 reduces the pu.1 signal, a marker for myeloid progenitors. The embryos were injected and processed as in panels D-F, but with an antisense probe for pu.1. (J-O) Knockdown of aggf1 expression results in a reduced number of granulocytes and macrophages during zebrafish early embryogenesis. (J-L) Knockdown of aggf1 expression reduces the mpo signal, a marker for granulocytes. Zebrafish embryos (1- to 2-cell stage) were injected with 2 ng of Std-MO (J), 2 ng of aggf1 MO1 (K), or 2 ng of aggf1 MO1 together with 100 pg of aggf1 mRNA (L) and used for WISH at 28 hpf with an antisense probe for mpo. Knockdown of aggf1 expression reduces the l-plastin signal at 28 hpf, a marker for macrophages (M-O). All embryos are shown in a lateral view with the anterior on the left.