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Fig. 2. Specificity of PG1 morpholinos. Two morpholinos for each Hox PG1 gene were injected into the left-hand side of the embryo and Krox20 (r3 and r5) and Engrailed-2 (midbrain/hindbrain boundary: * in A-I) expression analysed. The hindbrain region of early tailbud stage embryos (anterior to the top) is shown for non injected controls (NIC; A,D,G), Hoxa1 1st (B: 80%, n=15) and 2nd (C: 70%, n=10) morpholinos, Hoxb1 1st (E: 85%, n=13) and 2nd (F: 70%, n=20) morpholinos and Hoxd1 1st (H: 90%, n=11) and 2nd (I: 80%, n=15) morpholinos. Overexpression of Hoxd1 morpholino insensitive RNA (J) and rescue of Hoxd1 1st and 2nd morpholinos with Hoxd1 RNA is also shown (K,L). Dotted line indicates the midline. |
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Fig. 3. The triple PG1 knockdown phenotype is more severe than the double knockdowns. Morpholinos for each Hox PG1 gene were injected either in double (D: A1/B1 52%, n=25; E: B1/D1 48%, n=29; F: D1/A1 64%, n=33) or triple (C: A1/B1/D1 94%, n=35) combinations into the left-hand side of the embryo and Krox20 and Engrailed-2 (*) expression analysed. The hindbrain region of early tailbud stage embryos is shown, with anterior to the top. Non-injected controls (NIC; A), and embryos injected on the left-hand side of the embryo with control morpholino (B) are also shown. Arrowheads indicate neural crest expression. |
