Fig. 5: Epithelium permeability during pancreatic development and its impact on ductal morphogenesis.
a, A schematic showing the experimental and imaging setup for the pancreatic explant permeability assay. b, Live images of GFP–LifeAct pancreatic explants (top, E11.5; bottom, E15.5) treated with 10 kDa Dextran-647. The yellow asterisks indicate lumina detected via GFP–LifeAct. Scale bar, 50 µm. c, A quantification of normalized levels of 10 kDa Dextran-647 (luminal dextran signal to neighbouring cytoplasmic dextran signal ratio, at mid-plane) in the lumen. The data are presented as the mean ± standard deviation, pooled from three independent experiments (n = 5 E11.5, n = 4 E13.5, n = 7 E15.5 and n = 4 E17.5 explant samples). d, A schematic showing the experimental and imaging setup for E15.5 pancreatic explants under control and cCPE-647 treatment for permeability assays. e, The live images of membrane-tdTomato-expressing E15.5 pancreatic explants under control and cCPE-647 treatment, treated with 3–5 kDa Dextran-488. Scale bar, 100 µm. f, A quantification of normalized levels of 3–5 kDa Dextran-488 (ductal/luminal dextran signal to neighbouring cytoplasmic dextran signal ratio) in the duct/lumen. The data are presented as the mean ± standard deviation, pooled from three independent experiments (n = 5 control and n = 5 cCPE-647-treated E15.5 explant samples). g, A schematic showing the experimental and imaging setup for E15.5 pancreatic explants under control and cCPE-647 treatment. h, The maximum-projected immunofluorescence images of control and cCPE-647-treated E15.5 pancreatic explants stained with Mucin-1, after 2 days of culture. The yellow arrowheads indicate isolated ductal lumina. Scale bar, 50 µm. i, A quantification of the mean isolated ductal lumen density (number of isolated lumina per volumetric tile; x = 88.5 µm, y = 88.5 µm, z = 24 µm). The data are presented as the mean ± standard deviation, pooled from three independent experiments (n = 4 control and n = 5 cCPE-647-treated E15.5 explant samples). The P values were determined by a two-sided Mann–Whitney test for c, f and i. NS, not significant.
