Fig. 1: ephrinB2/Ror2 interaction regulates apical constriction.

a Schematic of screening for ephrinB2-binding partners and selected candidates from ephrinB2 pulldown assay. Scale bar, 500 μm. See also Supplementary Data 1. b Schematic of the transverse section through neural plate and RNA in situ hybridization (HCR) assay. Neural plate (yellow), PM; pre-somatic mesoderm (magenta), NC; notochord (blue), AR; archenteron roof (green). Scale bar, 50 μm. c Co-IP assay with Ror2 (1 ng) and ephrin-B type ligands (500 pg) at Stage 13 Xenopus embryos. d Schematic of ephrinB2 serial deletion mutants in C-terminus. The PDZ-binding motif in red. e Schematic of Ror2 serial deletion mutants in the tail region. f Co-IP with Ror2 (1 ng) and ephrinB2 (500 pg) serial deletion mutants. g Co-IP with ephrinB2 (500 pg) and Ror2 serial deletion mutants (1 ng). h Representative embryo morphology by ephrinB2 knockdown (2 ng of MO). See also Supplementary Movie 1. Schematic of D1.1 microinjection in the 16-cell stage embryo. Membrane-GFP indicates injected side (50 pg). The lateral boundaries of the neural plate are marked by dotted lines (green; uninjected side and magenta; injected side). Immunoblotting represents the expression level of ephrinB2 wildtype (WT) and Δ4 (25 pg). Data means SEM. Numbers of embryos/experiments: 85/3, 80/3, 78/3, and 79/3 from left to right for each column. One-way ANOVAs followed by Dunnett’s multiple comparison tests were used (p-value <0.0001). n.s., not significant, ****p < 0.0001. Scale bar, 500 μm. i Representative embryo morphology by Ror2 knockdown (2 ng of MO). See also Supplementary Movie 1. Schematic of D1.1 microinjection in the 16-cell stage embryo. Membrane-GFP indicates injected side. The lateral boundaries of the neural plate are marked by dotted lines (green; uninjected side and magenta; injected side). Immunoblotting represents expression levels of Ror2-WT, ΔFr, and ΔPR (50 pg). Data means SEM. Numbers of embryos/experiments: 79/3, 79/3, 81/3, 82/3, and 81/3 from left to right for each column. One-way ANOVAs followed by Dunnett’s multiple comparison tests were used (p-value <0.0001). n.s., not significant, ****p < 0.0001. Scale bar, 500 μm.. Scale bar, 500 μm.