IMAGE

FIGURE 4

ID
ZDB-IMAGE-211004-19
Source
Figures for Xia et al., 2021
Image
Figure Caption

FIGURE 4

Expression of human COG4p.G516R elevates the wnt4 transcript in zebrafish embryos. (A) Quantitative PCR analyses of selective non-canonical Wnt pathway ligands in zebrafish embryos injected with human COG4WT or COG4p.G516R mRNA. At 6 hpf, wnt4 expression is significantly upregulated in embryos injected with COG4p.G516R mRNA, but not in those injected with COG4WT mRNA. Similar upregulation of wnt4 transcripts is observed at 10 hpf as well. Bar graphs represent average gene expression relative to the housekeeping gene β-actin. The data are presented as mean ± SD. One-way ANOVA with Tukey’s multiple comparison tests was used. ****p < 0.0001; ns, not significant. (B) Representative images of whole-mount in situ hybridization from control and treated embryos. Whole-mount in situ hybridization analyses demonstrate that wnt4 expression is elevated in embryos injected with COG4p.G516R mRNA (a″, at 6 hpf; b″ at 12 hpf), but there are no obvious changes in embryos injected with human COG4WT mRNA (a′ and b′). (b, b’) Arrows point to the restricted wnt4 expression domain in the hindbrain of zebrafish embryos at 12 hpf; (b″) arrows point to expanded expression of wnt4 in the hindbrain region. At 12 hpf, wnt11f2 expression intensity is not significantly changed in embryos injected with COG4p.G516R mRNA (c″, d″), compared to either control siblings (c, d) or human COG4WT mRNA-injected embryos (c′, d′). The dotted black color lines landmark the degree of angle between the head and tail, which is much more increased in embryos injected with COG4p.G516R mRNA, suggesting defects in extension movement during gastrulation. Arrows in c, c′ and d, d″ indicate that wnt11f2 is restricted to the dorsal midline; however, its expression pattern is dispersed in COG4p.G516R mRNA-injected embryos, suggesting that the convergence movement is impaired (c″, white dotted line and arrow; d″, there is no clear midline expression). Scale bars: 200 μm. Experiments were performed in triplicates with similar results.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Front Cell Dev Biol