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. 2019 Apr 10;10:1659. doi: 10.1038/s41467-019-09655-3

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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. 6 — Functional interaction of mStau2 with oskar mRNA in Drosophila. a Expression of GFP-dmStau, GFP-mStau2, GFP-mStau2[F40A H169A] and GFP-mStau2[F40A F157A] in germline of stau^R9/stau^D3 mutant females. In dmStau-expressing oocytes oskar (cyan) localizes almost exclusively to the posterior pole (right) and bicoid (green) to the anterior pole (left) during stage 9 of oogenesis. Transgenic GFP-Stau protein is shown in red. In oocytes lacking Stau (stau null), oskar is found at both poles, enriching slightly more at the anterior, while bicoid localization is unaffected. Insets show magnified regions of the upper anterior corner. Scale bar: 20 µm and 1 µm for insets. b Typical localization of oskar mRNA in oocytes as function of expressed Stau protein. Using image transformation algorithms, RNA signal was redistributed into a 100 × 100 square matrix and statistically evaluated to obtain average (green) and variability (magenta) of RNA distribution. In wild-type oocytes (top left) most signal is found close to the posterior pole (right of the panels) by stage 9. In absence of Stau (stau null), oskar mRNA accumulates at the anterior pole. Scale bar: 20% length of anteroposterior axis. c, d Center of mass (relative to geometric center at 0, c) and fraction at posterior pole of oskar mRNA (d) during stage 9. P-values show result of pairwise Mann–Whitney U tests vs. the stau null condition (Bonferroni corrected alpha value: 0.01). N = number of oocytes. Center line: median; box limits: 25^th and 75^th percentile; whiskers: 10^th and 90^th percentile. e–h Interaction of GFP-tagged Stau molecules with oskar (e, f) and bicoid (g, h) mRNPs. mRNPs are sorted by their mRNA content using quantitative smFISH. Fraction of Stau positive mRNPs (e, g) and normalized GFP-Stau signal intensity (f, h) were plotted as function of mRNA content of the mRNPs. The normalized GFP-Stau signal intensities are fitted linear models, with indicated slopes. In pairwise comparisons of oskar mRNPs (f), all slopes are significantly different (p < 0.0001), except for GFP-mStau2[F40A H169A] vs GFP-mStau2[F40A F157A] (p = 0.016, alpha_corrected = 0.01). In bicoid mRNPs (h), the slope of GFP-mStau2[F40A F157A] differs from the other three (p < 0.01), which have similar slopes (p > 0.9). Source data are provided as a Source Data file