Expression of metap2 during embryonic development and metap2 knockdown with morpholino. (A) WISH showing the spatial expression of metap2 at different developmental stages. BA indicates brachial arches; ISR, intersomitic region. (B) Diagram of the action of the metap2 morpholino. Red line represents the binding site of the splice-junction targeted metap2 morpholino, and arrows represent the binding sites of the forward (P_f) and reverse primers (P_r) used for PCR amplification to detect defective splicing of exon4. (C) Defective splicing of metap2 mRNA in metap2^MO as shown by the presence of the smaller 557-bp band, which is absent in the control. Open arrowhead, 500-bp DNA ladder; filled arrowhead, 650-bp DNA ladder. (D) General morphology of metap2^MO (122 of 131 embryos in 3 separate experiments) compared with the control at 18 hpf (137/145, n = 3). (E-F) General morphology of metap2^MO (F; 109/129, n = 3) compared with the control (E; 132/142, n = 3) at 48 hpf. Bars represent 250 μm unless otherwise stated.