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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
Knockdown Reagent:
Anatomical Terms:
Stage: Long-pec
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
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