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. 2022 Oct 28;13:6423. doi: 10.1038/s41467-022-34165-0

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a Orthogonal (XZ) projections of myosin II recruitment to the MCC leading edge during integration. MCC is labeled with α-tubulin::LifeAct-GFP (pseudo-colored in green in the composite image, pseudo-colored in gray as a separate channel) and myosin-II is labeled with the myosin intrabody SF9-3xGFP (pseudo-colored in magenta in the composite image, fire pseudo-colored as a separate channel). White arrowheads depict myosin recruitment. Scale bar: 5 μm. b Normalized myosin-II intensity (magenta) and normalized F-actin intensity (green) at the cortex of integrating MCCs. T = 0 marks the last tracked frame during integration (n = 10 cells from 3 embryos, N = 3 experiments). Data show mean ± SEM. c Image sequence of control MCC (yellow arrowhead) and CA-MYPT-overexpressing MCC (white arrowhead, marked with H2B-RFP pseudo-colored in magenta) during integration. Control and CA-MYPT overexpressing MCC express LifeAct-GFP mRNA (pseudo-colored in green). Scale bar: 10 μm. d Quantification of MCC integration success rates for control and CA-MYPT overexpressing cells (nWT = 55 cells, nCA-MYPT = 24 cells from 5 embryos, N = 5 experiments). e Image sequence of a CAMYPT-overexpressing MCC attempting epithelial junction remodeling (orange arrowheads). Goblet cells and CA-MYPT overexpressing MCC express LifeAct-GFP mRNA (pseudo-colored in green). Scale bar: 5 μm. f Junction length tracking for control (green) and CA-MYPT-overexpressing MCCs (magenta) (nWT = 12 junctions, nCAMYPT = 13 junctions from 5 embryos, N = 5 experiments). Boxplots of final to initial junction length ratio in control and CA-MYPT MCCs. The orange line indicates no overall junctional remodeling (final to initial junction length ratio = 1). Boxes extend from the 25th to 75th percentiles, with a line at the median and whiskers representing range within 1.5 interquartile range. Two-tailed Mann–Whitney test with 0.05 significance level *p = 1.73E−5.