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. 2022 Jun 21;13(7):1107. doi: 10.3390/genes13071107

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Calcein staining illustrating the impact of foxl1, foxc1a, and foxc1b lone and combined loss on axial skeleton development and calcification. (A) WT, foxl1^-/-, foxc1a^-/-, and foxc1b^-/- embryos at 6 dpf showing normal formation of the vertebrae in foxl1 and foxc1b mutants. foxc1a^-/- embryos did not develop or calcify vertebrae and had large abdomen cavity edemas that grew over time. (B) WT; foxc1a^-/-; foxl1^-/-, foxc1b^-/-; foxl1^-/-, and foxc1a^-/-; foxc1b^-/- embryos at 6 dpf. Double foxc1a^-/-; foxl1^-/- embryos exhibit larger edemas and a lack of most calcified structures at 6 dpf, including the operculum (OP). foxc1b^-/-; foxl1^-/- embryos had no significant change. foxc1a^-/-; foxc1b^-/- mutants elicited similar results as foxc1a^-/-; foxl1^-/- embryos, including edemas and lack of craniofacial calcification. (C) Boxplots illustrating the variability in the number of vertebrae calcified in foxl1^-/-, foxc1a^-/-, foxc1b^-/-,and foxl1^-/-; foxc1b^-/- mutants in comparison with WT embryos at 6 dpf. Red arrows indicate primary centres of ossification within the vertebrae. Scale bars are 500 µm.