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. 2021 Mar 2;11:4976. doi: 10.1038/s41598-021-83771-3

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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In vitro migration of iPSC derived wild type and mutant cranial neural crest cells. (a) Representative images of scratched area for BJ1 wild type derived CNCC to migrate into at 0 h. Cells have been transfected with mRNA 24 h prior to imaging. From left to right: Bright field, cytosolic free-floating mClover3 and nuclei labelled with H2B-mCherry. Scale bar is 50 μm. Also see Additional Movie Files 1, 2 and 3. (b) Left: image of the same scratched area in (a) after 24 h of migration visualizing labeled nuclei with H2B-mCherry. Right: Representative image of tracking the migration path of individual neural crest cells using ImageJ plugin TrackMate. (c) Quantification of the neural crest migration speed. Cells carrying either the heterozygous or the homozygous Q152* mutation (Q152*^KI/WT, Q152*^KI/Q152*^KI) displayed a lower migration speed compared to their isogenic control line WT/WT. No difference between the full knock down Q152*^KI/Q152*^KI of MAPRE2 compared to the heterozygous knock down Q152*KI/WT was noted (One-Way ANOVA, Tukey test, P = 0.045, P = 0.0423 and P = 0.9996, respectively). In contrast, the N68S/N68S mutant NC cells migrated faster than their respective isogenic control line N68S^IC/N68S^IC (Unpaired t-test, P = 0.044). N = 11 for each genotype.