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. 2021 Aug 1;32(16):1501–1513. doi: 10.1091/mbc.E21-05-0258

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© 2021 van Loon et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 5: — Inhibiting NMII changes microridge patterns. (A) Representative images of periderm cells expressing Lifeact-GFP on 72 hpf zebrafish after 24-h exposure to the specified concentration of blebbistatin or vehicle control (DMSO). Images were inverted, so that high intensity fluorescence appears black and low intensity is white. (B) Visualizations of microridge distances, orientations, and alignment domains from periderm cells at 72 hpf, after 24-h exposure to the specified concentration of blebbistatin or vehicle control (DMSO). (C) Violin and box-and-whisker plot of projection length for periderm cells in 72 hpf zebrafish, after 24-h exposure to the specified concentration of blebbistatin or vehicle control (DMSO). DMSO, n = 26 cells from 9 fish; 5 μM blebbistatin, n = 27 cells from 9 fish; 50 μM blebbistatin, n = 29 cells from 9 fish. P < 2.2 × 10^–16, Kruskal–Wallis test followed by Dunn’s test with Benjamini-Hochberg p value adjustment: DMSO-5 μM blebbistatin, P = 0.173; DMSO-50 μM blebbistatin, P = 2.51 × 10^–13; 5 μM blebbistatin-50 μM blebbistatin, P = 3.79 × 10^–17. (D) Dot and box-and-whisker plot of microridge density, defined as the sum microridge length (μm) normalized to apical cell area (μm²), for periderm cells in 72 hpf zebrafish after 24-h exposure to the specified concentration of blebbistatin or vehicle control (DMSO). DMSO, n = 26 cells from 9 fish; 5 μM blebbistatin, n = 27 cells from 9 fish; 50 μM blebbistatin, n = 29 cells from 9 fish. P = 2.80 × 10^–14, one-way ANOVA followed by Tukey’s HSD test: DMSO-5 μM blebbistatin, P = 3.07 × 10^–3; DMSO-50 μM blebbistatin, P <2 × 10^–16; 5 μM blebbistatin-50 μM blebbistatin, P = 7.70 × 10^–8. (E) Dot and box-and-whisker plot of the mode distance between neighboring microridges in periderm cells in 72 hpf zebrafish after 24-h exposure to the specified concentration of blebbistatin or vehicle control (DMSO). DMSO, n = 26 cells from 9 fish; 5 μM blebbistatin, n = 27 cells from 9 fish; 50 μM blebbistatin, n = 29 cells from 9 fish. P = 0.318, one-way ANOVA. (F) Dot and box-and-whisker plot of the alignment index on periderm cells in 72 hpf zebrafish after 24-h exposure to the specified concentration of blebbistatin or vehicle control (DMSO). DMSO, n = 26 cells from 9 fish; 5 μM blebbistatin, n = 27 cells from 9 fish; 50 μM blebbistatin, n = 29 cells from 9 fish. P = 4.56 × 10^–7, one-way ANOVA followed by Tukey’s HSD test: DMSO-5 μM blebbistatin, P = 1.11 × 10^–5; DMSO-50 μM blebbistatin, P = 2.38 × 10^–6; 5 μM blebbistatin-50 μM blebbistatin, P = 0.951. Scale bars: 10 µm (A) and 5 μm (B). **p ≤ 0.01 and ***p ≤ 0.001. For box-and-whisker plots, the middle line is the median, and bottom and top ends of boxes are 25th and 75th percentiles, respectively.