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. 2023 Mar 15;13(3):220367. doi: 10.1098/rsob.220367

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© 2023 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 3. — Apical membrane localized Flotillin2 is required for notochord lumen formation. (a,b) ELMOD3 bound with Flotillin2. ELMOD3 interacted with Flotillin2 by Co-IP assay in HEK-293T cells and Y2H assay. (c) The overexpression of Flotillin2 mutants prevented lumen formation. Left panel: the Flotillin2 wild-type or mutant construct was overexpressed in notochord cells, respectively. Yellow asterisks indicate the notochord cell overexpressing Flotillin2 wild-type or mutants. Right panel: schematic representation of the phenotype of mutant overexpression. (d) The knockout of Flotillin2 suppressed lumen formation. Left panel: confocal images of Flotillin2-KO and control group embryos at 21 hpf, respectively. Yellow asterisks indicate the notochord cell of Flotillin2-KO. Right panel: schematic images depict the phenotypes of the Flotillin2-KO. (e) The apical localization of Flotillin2 depends on ELMOD3. Left panel: the co-expression of ELMOD3 wild-type or mutants and Flotillin2 in notochord cells and the localization of Flotillin2 is abnormal in these notochord cells overexpressing ELMOD3 mutants compared with control. Right panel: schematic diagrams depicting Flotillin2 localization. The grey, red and blue areas represent the nuclei, Flotillin2 and lumen, respectively. Scale bars, 10 µm.