Morpholino-mediated knockdown of zebrafish Emilin3 transcripts. (A) RT-PCR analysis of RNA extracted from 24 hpf non-injected embryos (WT) and embryos injected with the indicated morpholinos. β-actin was used as a loading control. (B) Whole mount immunofluorescence labeling for Emilin3 in 24 hpf embryos injected with 4 ng control morpholino (Ctrl MO), 2 ng/each of emilin3a and emilin3b splice-blocking morpholinos (e3a+e3b MO splice), or 2 ng/each of emilin3a and emilin3b translation-blocking morpholinos (e3a+e3b MO ATG). Scale bar, 50 μm. (C) Lateral views of 24 hpf embryos injected with 4 ng control morpholino (Ctrl MO) or 2 ng/each of emilin3a and emilin3b translation-blocking morpholinos (e3a+e3b MO ATG). Arrows point at notochord distortions in Emilin3 double morphants. (D) Lateral views of 24 hpf embryos injected with 10 ng control morpholino (Ctrl MO), 2 ng/each of emilin3a and emilin3b splice-blocking morpholinos (e3a+e3b MO), 6 ng p53 translation-blocking morpholino (p53 MO) and the combination of p53, emilin3a and emilin3b morpholinos (e3a+e3b+p53 MO). no, notochord.
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