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. 2019 Dec 17;10:5744. doi: 10.1038/s41467-019-13702-4

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© The Author(s) 2019

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. 4 — a Illustration of image subtraction step assuming parallel (left panel) or antiparallel filament bundles (right panel). For parallel bundles of treadmilling filaments, image subtraction results in fluorescent spots corresponding to FtsZ binding or detaching from a filament bundle allowing to visualize polymerization (growth, green) and depolymerization (shrinkage, magenta) trajectories. We expect that spots corresponding to growing and shrinking ends have similar intensities and closely follow each other. In contrast, for antiparallel bundles of treadmilling filaments, the spots corresponding to growth and shrinkage would move in opposite directions and cross each other. b Raw image of FtsZ filament network and the corresponding differential image overlaid showing growing (green) and shrinking (magenta) ends without ZapA and (c) in the presence of 6.0 μM ZapA. Orange arrows indicate treadmilling direction. Right: Intensity distribution of fluorescence spots generated from a differential time-lapse movie of FtsZ/FtsA filaments and with 6.0 μM ZapA (c). Insets depict the mean normalized intensities and standard deviation (shaded error bands) of spots versus trajectory length. The presence of ZapA does not change fluorescent spot intensity and accordingly filament orientation. Scale bars: large image, 5 μm; small image, 2 μm. Source data available at Figshare [10.6084/m9.figshare.10347062.v1].