FIGURE

Fig. S5

ID
ZDB-FIG-130618-38
Publication
Yao et al., 2013 - Anaplastic lymphoma kinase is required for neurogenesis in the developing central nervous system of zebrafish
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Fig. S5

alk MO efficiency test experiments. (A) A schematic diagram showing arrangement of the first four alk exon-intron boundaries and MO binding sites. Numbers show exon/intron sizes in basepairs. (B) A construct containing egfp inserted in frame with the alk ATG site was generated to test the binding efficiency of ATG-MO. (C–F) Injection of this mRNA in combination with MOs resulted in different EGFP translation levels. Embryos at 8 hpf, with bright field images on top and fluorescent images below. Injection of the mRNA resulted in EGFP signal (D). Co-injection with alk ATG-MO blocked its translation (E). alk ATGmismatch-MO did not block its translation (F). Embryos labeled with asterisks in (C) and (E) were taken from the group shown in (D), indicating sufficient fluorescent excitation. (G) A diagram showing aberrant splice products with retention of intron 3, when alk pre-mRNA splicing is blocked by splicing MOs. (H) RT-PCRs were used to test splicing MO efficiency by primers indicated in (A). In Spl-MO injected 24 hpf embryos, the PCR product size was increased by approximately 300 bp (2nd lane) when compared to uninjected wild-type (1st lane), indicating retention of intron 3. -RT controls (3rd and 4th lanes, without reverse transcriptase in cDNA synthesis) excluded contamination by genomic DNA. gapdh was used as loading control.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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