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Rasip1 shuttles between junctions and apical compartments in close association with Myosin. A Schematic drawing of recombined Rasip1-Scarlet-I and GFP-Rasip1. B Antibody staining for Rasip1 and Cdh5 in wild-type embryos at 32 hpf. Consistent observations were made across 14 samples from 4 independent experiments. C Expression of Rasip1-Scarlet-I and Cdh5-Venus in wild-type embryos at 32 hpf. In both (B) and (C), white arrowheads indicate Rasip1 clusters, while white arrows label linear Rasip1 localizations along the junctions. Consistent observations were made across 22 samples from 7 independent experiments. D Time-lapse images of Cdh5-Venus and Rasip1-Scarlet-I with Rasip1 enriched at constricting junctions (arrowheads). E Time-lapse images of Cdh5-Venus and Rasip1-Scarlet-I with Rasip1 enriched at the apical compartment (arrows). F GFP-Rasip1 relocating from the apical compartment to the constricting junctions. Lines label the position of the apical compartment before contraction. Arrowheads mark the GFP-Rasip1 enrichment at the constricting junctions. G GFP-Rasip1 relocating from junctions to apical compartments as clusters. Lines indicate GFP-Rasip1 localized near the anterior boundary of the apical compartment. Arrowheads mark the GFP-Rasip1 clusters within the apical compartment. Masks were applied to focus solely on the apical domain. Consistent observations were made in 15 samples from 7 independent experiments (D–G). H Time-lapse images of GFP-Podxl1 and Rasip1-Scarlet-I during constrictions. Arrowheads indicate highly enriched Rasip1 at the constricting sites. I, J Time-lapse images showing the local enrichment of Myl9a-GFP and Rasip1-Scarlet-I and subsequent depletion at apical compartments (I) or along junctions (J). Arrows mark clusters where Rasip1 and Myl9a overlap, while arrowheads indicate Myl9a-positive, Rasip1-negative clusters. Masks were applied to focus solely on the apical domain. Consistent observations were made in 8 samples from 3 independent experiments. All scale bars: 5 μm.
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