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FIGURE 5. Knockdown of Whsc1 and Tacc3 decrease CNC migration in vivo. (A,B,F,G,K,L,P,Q) Anterior lateral views of tailbud stage embryos (depicted at st. 27), following whole mount in situ hybridization against twist. Each column of panels (A,B,F,G,K,L,Q) are lateral views of two sides of the same embryo. (C–E,H–J,M–O,R–T) Measurements were taken for the total area of the three PA (Arch 1-3 extend anterior to posterior), the length of each individual arch, and the migration distance, as measured from the dorsal most tip of each arch to the neural tube. Embryos were stained and quantified at stages 25–30. (K–T) Letm1 or Whsc2 KD did not significantly affect any of the measured parameters. (F–J) Tacc3 KD expression caused an 8.33% decrease in the total PA area, but did not affect length or arch migration. (A–E) Whsc1 KD caused a 23.57% decrease in PA area. Additionally, the length of the second and third pharyngeal arches decreased by 14.72 and 31.70%, respectively. The migration distance of the first, second and third pharyngeal arches decreased by 15.75, 24.04, and 29.29%, respectively. Significance determined using a student’s paired t-test. (Embryos quantified: Whsc1 KD = 13, Tacc3 KD = 18, Whsc2 KD = 12, Letm1 KD = 19.) ∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗P < 0.05, n.s., not significant. Scalebar is 250 μm. Image published in: Mills A et al. (2019) Copyright © 2019 Mills, Bearce, Cella, Kim, Selig, Lee and Lowery. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Gene Synonyms Species Stage(s) Tissue twist1.L twist, twist1-a, twist1-b, X-twi, Xtwi, Xtwist X. laevis Throughout NF stage 27 cranial neural crest Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + nsd2 MO + NF27 (in situ hybridization) Fig. 5 B CDE Anatomical Phenotype abnormal cranial neural crest decreased cell migration in cranial neural crest Disease Wolf-Hirschhorn syndrome Xla Wt + tacc3 MO + NF27 (in situ hybridization) Fig. 5 G HIJ Anatomical Phenotype abnormal cranial neural crest decreased cell migration in cranial neural crest Disease Wolf-Hirschhorn syndrome Larger Image Printer Friendly View

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