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. 2019 Jul 1;9:9475. doi: 10.1038/s41598-019-45851-3

Figure 2.

Figure 2

Versican: the gene responsible for the lht phenotype. Genetic map of lht (A). Transcript linkage map place lht between Ol12338 and Ch28575. The numbers show the recombination events from 1,502 meiosis between the lht phenotype and linked genetic marker (A). Sequencing analysis of the genomic mutation in the versican 3′UTR. Arrow indicates the G-to-T transversion, arrowhead indicates the thymine insertion (B). Morpholino (MO) injection assay and quantification data (C,D). Versican MO was injected at the one-cell stage into embryos obtained from a WT cross. A morphant injected with versican MO (0.5 mM) phenocopies the lht mutant (C). Graphical representation of the dose-dependent effect of versican MO showing all the lht mutant phenotypes in each embryo (D). The results are represented as mean percentages of three independent experiments, Mean ±  SD. Western blot of versican and actin in WT and lht mutant embryos at 2 and 3 dpf (E). Immunohistochemistry staining for chondroitin sulfate at 3 dpf. The WT and lht mutant embryos were treated with chondroitinase ABC before incubation with primary antibodies (n > 10) (F). Arrow indicates chondroitin sulfate expression in the heart around the atrioventricular valve. Scale bar: 50 μm (F).