Fig. S6
- ID
- ZDB-FIG-180202-6
- Publication
- Joris et al., 2017 - Number of inadvertent RNA targets for morpholino knockdown in Danio rerio is largely underestimated: evidence from the study of Ser/Arg-rich splicing factors
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Injection of sMOsrsf5a in srsf5a-/- mutant embryos led to the previously observed morphant phenotype. A, No defects could be observed in WT and srsf5a mutant embryos at 48hpf. Injection of sMOsrsf5a in wt and mutant (obtained from homozygous parents) led to developmental defects, these were more severe in mutants. B, Analysis of srsf5a transcripts produced in CtrlMO and sMOsrsf5a injected wt or srsf5a-/- mutant embryos by RT-PCR. In srsf5a mutants, the level of wt transcript is very low, while the srsf5a intron3 form is stabilized (srsf5a-i3/153nt). C, Injection of srsf5a i3/153nt at 100pg or 200pg did not lead to any developmental defect in comparison to uninjected embryos. D, Protein production from the srsf5a-i3/153nt transcript was tested by injecting a flag tagged srsf5a-i3/153nt RNA (Srsfs5-i3/153nt-Flag) at one cell stage, followed by protein extraction and western blot analysis using the appropriate antibody. No protein production could be detected. A flag tagged srsf5a RNA (Srsf5a-Flag) was used as positive control. NI = protein extract from uninjected embryos. |