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. 2021 Aug 3;12:4680. doi: 10.1038/s41467-021-24852-9

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A Unilateral injection of increasing doses of SF3B2MO (10-30 ng) interferes with sox10 expression at the neurula stage, in a manner similar to SF3B4 knockdown. Injection of a control MO (CoMO) did not significantly affect sox10 expression. The SF3B2 knockdown phenotype is efficiently rescued by injection of human SF3B2 (HSF3B2; 10 pg or 100 pg) plasmid DNA. mRNA encoding the lineage tracer ß -galactosidase was co-injected with the MOs to identify the injected side (punctate red staining). The injected side is indicated by an asterisk. Dorsal view, anterior to top. The two rows show examples of the phenotype for each injection condition. Scale bar, 500 μm. B Quantification of the phenotypes. The numbers on the top of each bar indicate the number of embryos analyzed from seven independent experiments. C Unilateral injection of SF3B2MO (20 ng) reduces the expression of other neural crest genes at the neurula stage including tfap2e and snai2, and expanded the neural plate expression domain of sox2 (white brackets) on the injected side. sox9 was not affected in these embryos. The injected side is indicated by an asterisk. Dorsal view, anterior to top. Scale bar, 500 μm. D Quantification of the phenotypes. The numbers on the top of each bar indicate the number of embryos analyzed from three independent experiments. E At the tailbud stage, SF3B2MO (20 ng) injected embryos show a decrease in the length of the NC streams (arrows) as visualized by sox10 (stage 25), sox9, and twist1 (stage 28) expression. CoMO injection did not affect NC streams formation. Lateral view, dorsal to top, anterior to right. Scale bar, 250 μm. F Quantification of the phenotypes. The numbers on the top of each bar indicate the number of embryos analyzed from three independent experiments. G At stage 40, the expression of runx2 in the pharyngeal arches mesenchyme (arrows) is severely downregulated in SF3B2MO-injected tadpoles. The white line indicates the midline (frontal view). Scale bar, 250 μm. The quantification of the runx2 phenotype is shown in panel F. Source data are provided as a source data file.