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Fig. 5
Figure 5. Knocking down HuB using morpholino antisense oligonucleotide does not affect the migration or survival of the PGCs. Embryos of transgenic fish expressing EGFP-F.nanos 32UTR were injected at one-cell stage either with a control morpholino (ctr MO, A–D), or with a morpholino inhibiting hub mRNA translation (hub MO, E–H) and analyzed at 24hpf. HuB knockdown embryos show no severe deformations in somatic development (compare A and E) and properly clustered PGCs in the region of the developing gonad (compare arrowheads B and F). In control morpholino-injected (ctr MO) embryos, the HuB-GFP reporter signal is detectable in perinuclear granules (C), whereas no signal is detected in embryos treated with hub MO (G) demonstrating sufficient morpholino-mediated inhibition. Both morpholinos had no effect on the level and localization of a Vasa.DsRed protein expressed in the same embryos. I: RT-PCR for Hu family members performed on sorted PGCs and somatic cells from 7hpf embryos. Similar to hub (Fig. 2D), hur and hug are expressed in PGCs. Amplification of nanos and the use of 24hpf total embryo cDNA serve as controls.