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. 2017 Jul 7;28(14):1937–1949. doi: 10.1091/mbc.E17-01-0029

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© 2017 Wirshing and Cram. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 9: — Elevated myosin activity alters myosin organization and interaction with actomyosin bundles. (A) Confocal maximum intensity projections of fixed spermathecae from a WT animal and an animal treated with RNAi against myosin phosphatase, mel-11(RNAi), expressing moeABD::mCherry to label actin (red) and GFP::NMY-1 to label myosin (green). Note the homogeneous distribution of myosin in actomyosin bundles in WT and laterally associated myosin clusters that create transverse bands across several actomyosin bundles in a mel-11(RNAi) spermatheca. (B) Confocal maximum intensity projections from two 4D ovulation movies showing progression of mel-11(RNAi) phenotypes in selected cells. In movie 1, the actomyosin bundles rupture. The arrowhead indicates the site of bundle detachment. At 6:40, bundles are attached, and they retract by 10:00. By 23:20, the basal cell layer pulls back (last frame). In movie 2, misaligned actin bundles appear to be stabilized and reinforced (arrowhead). (C) Line-scan analysis along a single actomyosin bundle in movie 1 (indicated by a dashed line) at 5, 10, and 20 min after oocyte entry reveals that myosin clusters increase in intensity and grow by fusion of smaller clusters along a single bundle. In addition, actin fluorescence intensity is increased under prominent myosin peaks. Scale bar, 20 μm (A), 5 μm (B).