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. 2021 Oct 11;11:20111. doi: 10.1038/s41598-021-99521-4

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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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Observations of skeleton formation in the experimental starfish larvae. Morphology and skeleton formation were observed in control larvae (a1–a3), nonfeeding larvae (b1–b5), rapamycin (rapa)-treated larvae (c1–c5), and axitinib (axi)-treated larvae (d1–d5) at 7 dpf. (a1–d1, a2–d2) Living larvae. In the panel a2, the arrowhead indicates an adult skeletal rudiment and the dotted circle indicates aggregated mesenchyme cells. (a3–d3) Fluorescence images of larvae examined by immunohistochemistry using the mesenchyme-specific marker MC5. The green signal shows MC5 expression, whereas the red signal shows Chaetoceros calcitrans in the stomach or autofluorescence. At 7 pdf, the ratio of larvae with adult skeletons (b4–d4) and that of larvae with aggregated mesenchyme cells in the posterior dorsal region (b5–d5) were evaluated. Feeding larvae and DMSO-treated larvae were used as controls for nonfeeding larvae and inhibitor-treated larvae, respectively. Scale bars: 50 μm. M: mol/L.