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. 2020 Jun 5;11:2830. doi: 10.1038/s41467-020-16712-9

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© The Author(s) 2020

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. 5 — a The ability of the SPA-2L133A mutant to co-precipitate interacting proteins is compared to full-length SPA-2 (WT). The signal from mass spectrometry is compared to the WT precipitation according the scale shown in the legend. b CCP-1-mGFP precipitates SPA-2, but interacts weakly with the SPA-2 L133A mutant. Input and precipitated fractions are analyzed by western blotting for SPA-2. c Conservation, coiled-coil and coiled-coil dimer prediction for CCP-1. The calponin domain is boxed with a dashed line. The indicated regions of CCP-1 (r1-r3) were deleted using an mCherry-selectable marker fusion as described in Materials and Methods. Variant functionality is scored according to the color scale where 1 and 0 represent wild type and deletion mutant growth rates, respectively. Lower panels show localization of the indicated deletion variants. Dotted white lines show the hyphal outline. Scale bar = 10 μm. d CCP-1 tip-localization depends on SPZ-1, JNS-1 and SPA-2, but not LAH-2. The distribution of F-actin is shown for the indicated strains. Dotted white lines show the hyphal outline. Scale bar = 10 μm. e SPK F-actin signal intensity (arbitrary units) is quantified for wild type and ccp-1 deletion strains. Data are shown as mean values ± SD (n = 5 independent measurements shown as opaque red dots). The p-value calculated from 2-sided Student’s t-test is indicated. f LAH-2 depends on CCP-1, but other scaffold components do not. Localization of the indicated GFP-tagged proteins is shown in wild type and ccp-1 deletion mutant. Dotted white lines show the hyphal outline. Scale bar = 10 μm. Source data are provided as a Source Data file.