Overexpression of fli-vp16 or etsrp rescues defects in specification of hemangioblasts and hematopoietic cells in aggf1 morphants. (A-I) Zebrafish embryos (1- to 2-cell stage) were injected with Std-MO (4 ng; A,D,G), aggf1 MO2 (4 ng; B,E,H), or aggf1 MO2 (4 ng) together with fli-vp16 mRNA (75 pg) (C,F,I) and used for WISH at 10 somites with antisense probes for scl (A-C), lmo2 (D-F), and gata2 (G-I). Reduced expression of hemangioblast markers (scl, lmo2) in PLPM by aggf1 MO2 was restored by overexpression of fli-vp16 mRNA (A-F). Similarly, reduced expression of hematopoietic marker gata2 in PLPM by knockdown of aggf1 expression was restored by overexpression of fli-vp16 mRNA (G-I). (J-O) Zebrafish embryos (1- to 2-cell stage) were injected with Std-MO (2 ng; J,M), aggf1 MO1 (2 ng; K,N), or aggf1 MO1 (2 ng) together with etsrp mRNA (100 pg) (L, O) and used for WISH at 6 somites with antisense probes for scl (J-L) and lmo2 (M-O). Reduced expression of scl and lmo2 in PLPM by aggf1 MO1 was restored by overexpression of etsrp mRNA, suggesting that etsrp is downstream of aggf1 during differentiation of hemangioblasts. Embryos are shown in a dorsal view with the anterior at the top.