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Rasip1 inhibits contractility at junctional patches and apical compartments. A Antibody staining for Myl9a-GFP and Rasip1 at junctional patches and rings of varying sizes. Consistent observations in 10 samples from 3 independent experiments. B The Dice coefficient and corresponding area of various junctional patches and rings, with data points color-coded by area (n = 27 cells from 10 embryos). C Time-lapse imaging of Rasip1-scarlet-I and Myl9a-GFP in wild-type embryos during the patch-to-ring transitions. White arrows mark colocalized Myl9a and Rasip1 at the junctional patch, while white arrowheads indicate segregation at later time points. (C’) Automatic thresholding and respective graphical representations. Consistent observations in 6 samples from 3 independent experiments. D, E Cdh5-Venus and Myl9a-GFP in wild-type embryos (D) and rasip1 mutants (E). White arrows indicate Myl9a within the patches, while arrowheads mark Myl9a at the boundary. D’, E’ Automatic thresholding and respective graphical representations. Consistent observations in 10 samples from 5 independent experiments per group. F, G Distinct localizations of Myl9a-GFP in wild-type embryos (F) and rasip1 mutants (G). Arrowheads label Myl9a at junctions, while arrows indicate Myl9a clusters within the apical compartments. F’, G’ Intensities of signals along the dashed lines in (F, G). H Normalized Myl9a intensities at the peripheral regions and apical compartments in wild-type embryos and rasip1 mutants (WT: n = 25 cells from 6 embryos; rasip1: n = 18 cells from 8 embryos), presented as mean ± SD. The Myl9a-GFP intensity is standardized using the mean signal level from the entire cell. ***P = 0.0001, n.s., not significant, 2-tailed unpaired t-test. I–K Time-lapse images of Cdh5-Venus and Myl9a-GFP showing synchronized clustering in wild-type embryos (I) and in rasip1 mutants (J, K). Arrowheads indicate Cdh5 and Myl9a along the junctions, while arrows indicate the Cdh5 and Myl9a clusters within the apical compartment. (I’) Diagrams illustrate transient Cdh5 aggregation under Myl9a enrichment along the junctions in wild-type embryos. (J’, K’) Diagrams illustrate Cdh5 aggregation under Myl9a enrichment towards the apical compartments (J’) or persistent contraction along junctions in rasip1 mutants (K’). All scale bars: 5 μm. Source data are provided as a Source Data file.
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