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Fig. 5 | Biological Research

Fig. 5

From: DYRK1-mediated phosphorylation of endocytic components is required for extracellular lumen expansion in ascidian notochord

Fig. 5

Clathrin-mediated endocytosis (CME) is required for expansion in Ciona notochord. AB Both clathrin and dynamin 1, the main components of endocytosis, are localized at the center of the anterior and posterior membrane at the initiation of lumen formation. Both were mainly enriched in the apical membrane during lumen expansion. C Schematic representation of the mutant constructs of dynamin1 (S728A). DE Overexpression of dynamin1 (S728A) arrested lumen expansion significantly. The white circle depicts the edge of the lumen, and the white asterisk indicates lumen with phenotype, and the red asterisk indicates control lumen. “n” represents the statistical number of lumens. Asterisks (***) represent statistical significance (p < 0.001). F Treatment with endocytosis inhibitors (Chlorpromazine, 10 μM; Pitstop2, 5 μM; Dynasore, 80 μM) arrested lumen expansion. G Exocytosis markers present in the apical membrane during notochord lumen expansion. pHmScarlet, a fluorescent protein, fused with VAMP2. The result showed VAMP2::pHmScarlet localized at the notochord apical membrane at 23 hpf. HI Rab11A, the apical secreted vesicle coat protein, was expressed in notochord (promoter assay) H and localized at the apical membrane during notochord lumen expansion (I). Bars in A–B, D, F–G, and I are 20 μm. The bar in H is 100 μm

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