Fig. 3

Morpholino knockdown of Pitx3 protein and the morphant phenotypes. (A) Immunoblot analysis of protein extracts from Morph1-injected embryos (M), a five base mismatch control (mmC) and an uninjected control (uiC) at 3 dpf (lanes 2–4), 4 dpf (lanes 5–7), 5 dpf (lanes 8–10) and 7 dpf (lanes 11–13). Lane 1 reveals the 32 kDa Pitx3 protein in 24 hpf whole embryo extract using the anti-Pitx3 polyclonal serum. Pitx3 protein expression was reduced in the morphant extracts compared to the mismatch and uninjected controls. Immunodetection of actin revealed that equivalent amounts of protein were loaded for all three samples from each time point (not shown). (B–Q) The pitx3 morphant phenotypes. Lateral views of wild type, morphants and mismatch control fish at 3 dpf (B–E). The small morphant eyes are shown in lateral views relative to the wild type and mismatch controls at 3 and 5 dpf (F–H and I–K, respectively). At 5 dpf, the morphants lack normal lower jaws (J; arrow) and pectoral fins (M; arrow). At 7 dpf, the morphant eyes are further reduced in size compared to the wild type and mismatch control (O–Q).