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. 2021 Sep 1;12:5222. doi: 10.1038/s41467-021-25461-2

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© The Author(s) 2021, corrected publication 2021

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. 2 — A Scanning electron micrographs showing sporulating hyphae and spores of the wild-type and hupS, smc and hupS smc double mutant (ftsZ-ypet derivatives, MD100, TM005, TM004 and TM006, respectively) (representative of 10 images). Elongated spores are marked with a yellow arrowhead. Scale bar: 10 µm. B Box plot analysis of the spore length distribution in the S. venezuelae wild-type (light green), hupS (yellow), smc (blue) and hupS smc (pink) (AK200, TM010, TM003, respectively), as well as in hupS and smc mutants complemented with in trans-delivered hupS-flag (orange) and smc-flag genes (dark blue) (TM015 and TM019, respectively) (300 spores were analysed for each strain). Boxplots show median with first and third quartile while the lower and upper ‘whiskers’ extend to the value no further than 1.5 * IQR (interquartile range) from the ‘hinge’. The statistical significance between strains determined by a one-way anova with a Games–Howell post-hoc test (two-sided) is marked with asterisks: p-value ≤0.05 (*), ≤0.01 (**) and ≤0.001 (***). p-values: wild-type-hupS: <0.0001, wild-type-smc: 0.0242, wild-type-hupS smc: <0.0001, wild-type-hupS; hupS-FLAG: 0.0625, wild-type-smc; smc-FLAG: 0.3871, hupS-smc: 0.0089, hupS-hupS smc: 0.1241, smc-hupS smc: <0.0001. C Examples of time-lapse images (representative of 10 repetitions) showing visualisation of nucleoids (marked with HupA-mCherry fusion) and Z-rings (visualised by FtsZ-YPet fusion) in the wild-type background (ftsZ-ypet, hupA-mCherry derivative, TM011) hupS, smc and hupS smc double mutant background (ftsZ-ypet, hupA-mCherry derivative, TM013, TM012 and TM014, respectively). An abnormal hyphal fragment is marked with a yellow arrowhead. Scale bar: 5 µm. D Box plot analysis of the nucleoid area (visualised by HupA-mCherry fusion) in the wild-type (light green, 262 spores), hupS (yellow, 232 spores), smc (blue, 220 spores) and hupS smc (pink, 218 spores) double mutant (ftsZ-ypet, hupA-mcherry derivative, MD100, TM011, TM013, TM012, TM014, respectively). Boxplots show median with first and third quartile while the lower and upper ‘whiskers’ extend to the value no further than 1.5 * IQR (interquartile range) from the ‘hinge’. The statistical significance between strains determined by Wilcoxon rank-sum test (two-sided) with Holm method used for multiple comparisons is marked with asterisks: p-value ≤0.05 (*), ≤0.01 (**) and ≤0.001 (***). p-values: wild-type-hupS: <2e−16, wild-type-smc: <2e−16, wild-type-hupS smc: <2e−16, hupS-smc: 0.9082, hupS-hupS smc: 0.0028, smc-hupS smc: 0.0039. E Kymographs showing the intensity of Z-ring fluorescence along the representative hyphae in the wild-type and hupS, smc and hupS smc double mutant (ftsZ-ypet derivatives, MD100, TM005, TM004, TM006, respectively) during maturation. F The variation (standard deviation) of FtsZ-Ypet (Z-rings) fluorescence during spore maturation of strains calculated for 25–30 hyphae of the wild-type (light green), hupS (yellow), smc (blue) and hupS smc (pink) double mutant (ftsZ-ypet derivatives, TM005, TM004, TM006, respectively). Fluorescence was measured starting from 30 min before growth arrest until spore formation. Points represent values for each hyphae, while lines show the result of Local Polynomial Regression Fitting (loess) with 95% confidence intervals (shaded area).