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Apical clearance and lumenal defects in rasip1 mutants. A Antibody staining for Cdh5 and ZO-1 in wild-type embryos and rasip1 mutants at 32 hpf. White arrows indicate discontinuous junctions. White and green arrowheads indicate junctional clusters and linear junctional structures within the presumed apical compartments, respectively. B Expression of Cdh5-Venus in wild-type embryos and rasip1 mutants at 32 hpf. Green and magenta masks label the boundary regions (junctions) and the presumed apical compartments, respectively. C Quantification of the ratio of the average Cdh5 intensity between the boundary regions (green masks in (B)) and the apical compartments (magenta masks in (B)), hereafter referred to as the boundary-to-apical (B/A) ratio. (WT: n = 53 cells from 10 embryos; rasip1: n = 55 cells from 11 embryos). Data are presented as mean ± SD. ***P < 0.0001, 2-tailed unpaired t-test. D–F Cdh5-Venus and GFP-Podxl1 in wild-type embryos (D) and rasip1 mutants (E, F). (D’–F’) Intensities of Cdh5-Venus and GFP-Podxl1 along the dashed lines in (D–F). G Coefficient of variation of Podxl1 per apical compartment, presented as mean ± SD (WT: n = 13 cells from 6 embryos; rasip1: n = 14 cells from 7 embryos). ***P < 0.0001, 2-tailed unpaired t-test. H, I Time-lapse images of Cdh5-Venus and GFP-Podxl1 in wild-type embryos (H) and rasip1 mutants (I). White arrows in (I) indicates GFP-Podxl1 that emerged outside of junctions. Consistent observations in over 10 samples from 4 independent experiments per group. All scale bars: 10 μm. Source data are provided as a Source Data file.
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