Castro et al., 2020 - Activation of WNT signaling restores the facial deficits in a zebrafish with defects in cholesterol metabolism. Genesis (New York, N.Y. : 2000)   58(12):e23397 Full text @ Genesis

Fig. 1 Inhibition of cholesterol synthesis is associated with decreased expression of WNT target genes. (a) RNA was isolated from homozygous carriers of the Vu57 allele (Vu57) or wild type siblings (Sibling) at 30 hours post fertilization (HPF) and the relative expression of axin2, edn1, lef1, and ccnd1 were measured by quantitative real time PCR (qPCR). N = 8/group in two biological replicates for a total N of 16. *p < .05. (b) Wild type larvae were treated with vehicle control (DMSO), 2 μM atorvastatin (ATOR), 8 μM lonafarnib (lona), or 1.5 μM Ro‐48‐8071 at sphere stage. Total RNA was isolated from a pool of embryos (N = 14) and the relative expression of axin2 was measured by quantitative real time PCR (qPCR). (c) Embryos were treated with vehicle control (DMSO), 1, 2, or 3 μM Ro 48‐8071 at the sphere stage and total RNA was isolated at 30 HPF from a pool of embryos (N = 30) and the relative expression of axin2 was measured by qPCR. *p < .05. Error bars represent SD. (d). RNA was isolated from homozygous carriers of the Vu57 allele or Sibling wild type at 30 HPF and the relative expression of dlx2a was measured using qPCR. N = 8/group with two biological replicates for a total N = 16. Error bars represent SD. (e) sox10 expression was monitored in embryos treated with vehicle control (DMSO), 1, 2, or 3 μM Ro 48‐8071 at the 30 HPF (N = 30). Error bars represent SD. *p < .05

Fig. 2 Activation of WNT signaling restores the facial defects present in the Vu57 allele. (a) Experimental design schematic with onset of treatment with WNT‐Agonist I at 30 hours post fertilization (HPF) and removal of treatment at 54 HPF. Alcian Blue was performed at 4 days post fertilization (DPF) in treated and untreated individuals that were incubated in embryo media from 54–96 HPF following treatment. (b–e) Alcian blue staining was performed at 4 DPF in homozygous carriers of the Vu57 allele (Vu57−/−) and wild type siblings (Sibling) according to the treatment schematic in (a). Total numbers of animals reflected in Table 1. *p = .0001 using a Fisher's Exact test. (f). Homozygous carriers of the Vu57 allele (Vu57−/−) or wild type siblings (Sibling) were treated with vehicle control (DMSO) or WNT‐agonist I at either 0.1 or 0.2 μM concentration according to the schematic in (a). Total RNA was isolated from a pool of embryos and the expression of sox10, col2a1a, or axin2 was measured by quantitative PCR (qPCR). Error bars represent SD of biological replicates. N = 11/group except for 0.2 μM concentration where N = 9. *p = 4.07457E‐05, **1.14579E‐06, ***p = 3.80572E‐07, #2.45716E‐05, ##p = 7.28315E‐08, ###p = .000716073, &p = 3.22846E‐05, &&p = 7.2866E‐07, &&&p = 8.45279E‐07

Acknowledgments:
ZFIN wishes to thank the journal Genesis (New York, N.Y. : 2000) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Genesis