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etv7 morphants express beta-globin (hbbe1.1). (A) In situ hybridization analyzing the expression of beta-globin mRNA was performed on embryos injected with 8.2 ng of standard control morpholino (Std) or 8.2 ng of translational-blocking morpholino (MT ATG) at 23, 24, 33 or 48 hpf. Arrows indicate expression in the posterior blood island (PBI) and arrowheads depict expression on the yolk sac. (B) Semi-quantitative PCR of beta-globin (hbbe1.1) expression at 24 hpf. actin expression is shown as loading control. Std are animals injected with 8.2 ng of standard control morpholino, MT ATG are animals injected with 8.2 ng of translational blocking morpholino and Neg is control PCR reaction without template. (C) etv7 morphants exhibit reduced levels of mature red blood cells. o-dianisidine staining was performed at 30 hpf with embryos injected with 8.2 ng of translational-blocking morpholino (MT ATG) or 8.2 ng standard control morpholinos (Std).
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