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ZIRC
ZFIN ID: ZDB-FIG-140512-20
Osborn et al., 2014 - Loss of FTO antagonises Wnt signaling and leads to developmental defects associated with ciliopathies. PLoS One   9(2):e87662 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Long-pec

Fig. 2

Canonical Wnt signaling is downregulated in fto morphants zebrafish.

(A) Dual luciferase assay using the β-catenin responsive TopFlash construct shows loss of reporter assay activity in ftoMO embryos analysed at both 24 hpf (con: 1.000 SEM ±0.073, ftoMO: 0.491 SEM ±0.105) and 48 hpf (con:1.000 SEM ±0.103, ftoMO 0.580 SEM ±0.066) stages. (B) Lef1 transcripts, a canonical Wnt target gene, were analysed by in situ hybridisation (ISH) at 48 hpf. Fto morphants showed marked loss of lef1 expression in the optic-tectum (arrows). Scale bar: 500 μm. n = con 56/66, ftoMO 40/53. (C) Loss of β-Catenin was confirmed in fto morphants by western blotting at 48 hpf. β-Catenin protein levels were quantified relative to the loading control (Actin). (D) ISH analysis of ctnnb1 (zebrafish β-catenin 1) at 48 hpf showed upregulation of transcripts specifically in areas of the lateral hindbrain (arrowheads). Scale bar: 500 μm. n = con 70/70, ftoMO 65/68 (E) Fto morphant Tg(7xTCF-Xla.Siam:GFP)ia4 display loss of GFP accumulation in both the Telen- and Diencephalic regions of the brain when compared to uninjected controls at 48 hpf, embryos viewed from a dorsal perspective. Scale bar: 100 µm; n = con 20/20, ftoMO 18/20.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ctnnb1 AB/TL control Long-pec hindbrain ISH
AB/TL + MO1-fto standard conditions Long-pec hindbrain ISH
GFP ia4Tg control Long-pec diencephalon IFL
Long-pec telencephalon IFL
ia4Tg + MO1-fto standard conditions Long-pec diencephalon IFL
Long-pec telencephalon IFL
lef1 AB/TL control Long-pec optic tectum ISH
AB/TL + MO1-fto standard conditions Long-pec optic tectum ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB/TL + MO1-fto standard conditions Long-pec canonical Wnt signaling pathway disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One