Figure 3

GFP expression in GATA-2 transgenic zebrafish. (A) Whole mount in situ hybridization shows GATA-2 mRNA expressed in zebrafish embryonic brain, ganglia cells, motor neurons and hematopoietic ICM region at 24 hpf stage. Arrows indicate the ICM. (B) Transgenic zebrafish carrying a modified BAC construct containing a 20 kb genomic sequence upstream of, and adjacent to the GATA-2 start codon show GFP expression in the ICM, brain and spinal cord at 24 hpf stage. (C) Transgenic zebrafish embryos carrying a GFP reporter gene construct linked to a 7.3 kb genomic sequence upstream of and adjacent to the GATA-2 start codon have strong GFP expression in neuronal cells, but not in the ICM at 24 hpf. (D) Transgenic zebrafish embryos carrying BACΔUp1/GFP have strong GFP expression in the ICM at 24 hpf stage. (E) Transgenic zebrafish embryos carrying BACΔUp2/GFP and (F) BACΔUp1ΔUp2/GFP have GFP expression in neuronal tissue but not in the ICM. (G) Transgenic zebrafish embryos carrying Up1-mp/GFP have strong GFP expression in the heart at 24 phf stage. (H) Stable transgenic zebrafish embryos containing the Up2 element linked with mp/GFP have strong GFP expression in the ICM and weak GFP expression in the spinal cord at 24 hpf stage. (I) Transgenic zebrafish embryos carrying Up2a-mp/GFP have GFP expression in ICM region at 24 hpf stage; non-specific GFP expression was observed the in notochord. (J-L) Transgenic zebrafish embryos carrying base change mutation in Up2 element derived from construct Up2-mp/GFP. (J) Mutation in the binding site of ARP1 shows no effect on the GFP hematopoietic expression. (K) Mutation in the binding site of E2F-1 results in weaker GFP expression in ICM and strong non-specific expression in muscle and notochord. (L) Mutation in HOXA3-A binding site shows significantly reduced GFP expression in ICM. Similar reduction of GFP in ICM was seen in LMO2 mutation (see Additional file 3, Supplementary Figure S2). White arrows indicate ICM expression of GFP.