|
Impaired primitive erythropoiesis caused by <italic>cul4a</italic> knockdown is mediated by downregulated expression of <italic>scl</italic>.a The relative expression of the two isoforms of scl, scl-α and scl-β, were measured by qRT-PCR in embryos injected with CoMO or cul4a-MO. bo-dianisidine staining showed depletion of erythroid cells in cul4a-morphant embryos compared with controls. Zebrafish scl-α, but not scl-β, mRNA rescued the reduction in primitive erythropoiesis of cul4a-morphant embryos. c, d WISH was performed with gata1 or hbbe3 probes in 24 hpf embryos injected with CoMO or cul4a-MO, or MO coinjected with zebrafish scl-α or scl-β mRNA. e, f Relative expression of gata1 or hbbe3 in embryos injected with CoMO or cul4a-MO, or MO coinjected with zebrafish scl-α or scl-β mRNA at 24 hpf. g WISH was performed with lmo2 probes in embryos at 6 somite and 24 hpf, respectively. Reduced expression of lmo2 in cul4a-morphant embryos were rescued by coinjection of zebrafish scl-α, but not scl-β mRNA. h Relative expression of lmo2 was measured by qRT-PCR in embryos injected with CoMO or cul4a-MO, or MO coinjected with zebrafish scl-α or scl-β mRNA at 6 somite and 24 hpf. The number in the top right-hand corner indicates the phenotypic embryos/total embryos. qRT-PCR experiments were performed in triplicate. ***p < 0.001. All scale bars represent 250 μm
|
