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. 2020 Jul 23;11:3698. doi: 10.1038/s41467-020-17452-6

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© The Author(s) 2020, corrected publication 2022

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. 2 — a Representative bright field lateral images of WT and fam50a KO are shown at 3 and 5 days post-fertilization (dpf). Morphology of fam50a KO was relatively normal at 3 dpf; repeated four times. However, at 5 dpf, fam50a KO showed craniofacial abnormalities; repeated five times. Number of larvae assessed with similar results: 3 dpf, n = 56; 5 dpf, n = 44. b Fluorescent lateral images of WT and fam50a KO larvae on a Tg(huc:egfp) neuronal reporter. No difference between WT and fam50a KO was detected in the anterior structures at 2 dpf. However, at 3 dpf, a prominent reduction of GFP-positive neurons was observed in KO larvae. Number of larvae assessed with similar results: 2 dpf, n = 20; 3 dpf, n = 20. c Whole-mount in situ hybridization (WISH) of her4.1, a molecular marker of neurogenesis, indicated a depletion of neurons in fam50a KO compared to WT at 3 dpf; repeated. Number of larvae assessed with similar results: 2 dpf, n = 24; 3 dpf, n = 21. d Apoptosis markers are elevated in fam50a KO larvae at 2 dpf. Note the induction of tp53 and tp53 target genes, mdm2 and cdkn1a (p21) in the cell proliferative zone in the midbrain region fam50a KO larvae at 2 dpf; repeated. Number of larvae assessed with similar results: tp53, n = 41; mdm2, n = 52; cdkn1a, n = 39. e Representative ventral images of Alcian blue staining of cartilage structures shows severe defects in cartilage development that become apparent at 3 dpf. Meckel’s cartilage, mc; palatoquadrate, pq; ceratohyal arch, ch; and ceratobranchial arches, cb. Number of larvae assessed with similar results: 2.5 dpf, n = 28; 3 dpf, n = 21; 4.5 dpf, n = 14. For all images: anterior, left; posterior, right. Scale bars; 200 μm.