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Efficacy and sepcificity of lhx5 morpholinos. (A-D) lhx5 morpholino knockdown alters pax6a and pax2a expression patterns. Embryos were dissected and flat mounted in glycerol after in situ hybridization. Dorsal views, rostral to the top. (E-G) lhx5 translation-blocking morpholino reduces lhx5-GFP mRNA translation. GFP fluorescence in the cell nucleus is reduced when 2 ng (F) or 4 ng (G) of the translational morpholino is injected into zebrafish embryos. (H) lhx5-splicing morpholino reduces wild-type lhx5 transcripts. Arrow indicates wild-type lhx5 or odc1 transcripts. Band above the wild-type lhx5 transcript is caused by splicing with a cryptic GT located in lhx5 intron 3 downstream from the canonical GT site at the exon 3/intron 3 junction; it results in a frame shift. Amplifications of odc1 transcripts serve as controls.
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