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. 2019 Jan 21;10:268. doi: 10.1038/s41467-018-08218-2

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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. 1 — S. japonicus maintains cellular aspect ratio over a range of volumes. a S. japonicus analog-sensitive wee1-as8 cells incubated with methanol (solvent control) or 20 μM ATP analog 3-BrB-PP1. Note the morphological transition in 3-BrB-PP1-treated cells occurring at a 4-h time point. b Wee1 inhibition initially causes differences in the diameters of two daughter cells (orange circles indicate cells treated with 3-BrB-PP1 for 2 h; gray circles represent solvent control). Shown are scatter plots, where either “thinner” (top left) or “thicker” (top right) daughter cell diameter measurements are on x-axis, and the ratios between diameters of the daughters are on y-axis. Graph (bottom right) compares cell diameter changes of the two daughters between control and 3-BrB-PP1-treated wee1-as8 and wild type populations. c Quantifications of cell length, width and aspect ratio at division of wee1-as8 cells shown in (a) and similarly treated wild type cells shown in Supplementary Fig. 1c. d Wee1-inhibited cells recover their original dimensions following the removal of the ATP analog from the growth medium. Shown are wee1-as8 cells treated with 20 μM 3-BrB-PP1 for 7 h, following the washout of the drug for 2 and 6 h. e Quantifications of cell length, width and aspect ratio at division of cells shown in (d). Methanol-treated wee1-as8 cells washed with growth medium (5th column) were used to control for cell number increase and washing procedure. f wee1-G788E mutant cells cultured at 24 °C (left) and 30 °C (right) overnight. g Measurements of cellular length, width and aspect ratio of the wild type as compared to wee1-G788E cells shown in (f). h S. japonicus cells after the shift from YE to EMM for 0, 4, 7 h and overnight. i Quantifications of cell length, width and aspect ratio at division before and after shift from YE to EMM overnight, for cells shown in (h). a, d, f, h Shown are single z-plane bright-field micrographs of live cells; scale bars represent 5 μm. b, c, e, g, i Quantifications presented as box plots with whiskers calculated by the Tukey method; n indicated in figures; p values derived from Kolmogorov–Smirnov test