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. 2021 Sep 14;9:720688. doi: 10.3389/fcell.2021.720688

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Copyright © 2021 Xia, Zeng, Tambe, Ng, Dong and Freeze.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Overexpression of wnt4 phenocopies zebrafish embryos injected with the COG4^p.G516G mRNA. (A) Graphs show the measured body length of each group after wnt4 mRNA injection at 3 and 7 dpf. The data are presented as mean ± SD. An unpaired two-tailed t-test was used. ^****p < 0.0001; ^∗∗p < 0.01; ^∗p < 0.05; ns, not significant. (B) Overexpression of wnt4 causes abnormal chondrocyte stacking and intercalation at Meckel’s cartilage. Ventral view of representative Meckel’s cartilage of zebrafish larvae after wnt4 injection at 4 dpf following WGA staining and imaged by a confocal microscope. (C) Overexpression of wnt4 causes cyclopia as expression of the COG4^p.G516G variant. Dorsal view of representative images of cyclopia compared to control. Two hundred picograms of wnt4 or COG4^p.G516G mRNA was used per embryo. Experiments were performed in triplicates with similar results.