Ductal cell rearrangement and lumen resolution require EphrinB1 and EphB3b. a–d Overview of the forming HPD epithelium by maximum projection of confocal stacks stained for PanCadherin (magenta) aPKC (white) in control and ephrinb1 and ephb3b mutants. Blue dashed squares indicate magnified CBD containing areas. a At 48 hpf, PanCadherin shows multiple cell layers in CBD (e.g., colored cells 1–3). b By 60 hpf, the CBD is largely a single-layered epithelium with apical domains facing the central lumen. c, dephrinb1 and ephb3b CBDs show disrupted epithelial organization, including luminal loops, with cells with multiple or extended apical domains (yellow arrowheads), and teardrop shaped-cells with reduced apical domains (white arrowhead). e Quantification of the longest continuous aPKC staining as a measure for CBD lumen elongation in controls, ephrinb1 and ephb3b mutants between 52 and 60 hpf. f Basal/apical membrane ratio was used to quantify CBD cell shapes in control (n = 26 cells), ephrinb1 (n = 28) and ephb3b (n = 28) embryos at 60 hpf. g Quantification of CBD cell height (distance between apical and basal membranes) in control (n = 26 cells), ephrinb1 (n = 28) and ephb3b (n = 28) embryos at 60 hpf. Scale bars: a = 5 μm, b–d = 20 μm. Statistical test: e, g = Student’s t-test, f = Fisher Exact. Errors bar show SEM; **p < 0,01, ***p < 0,001. Source data are provided as a Source Data file
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