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. 2019 Oct 22;10:4797. doi: 10.1038/s41467-019-12745-x

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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. 5 — Cell size distribution in silenced and control pIZ-plasmid cultures. a Left: phase contrast micrographs of Ctrl control, SB3×6 and SB2 slaB-silenced (left) and complemented cultures (-CC) (right). Scale bar: 10 µm. Right upper panel: scanning electron micrographs (gold coated) of slaB-silenced SB3×6 and Ctrl; scale bar: 5 µm (magnification × 2700). Right lower panel: Transmission Electron micrographs (magnification: × 6000) of thin sections (contrasted with Uranyl acetate 2%, and Lead citrate 2%) of slaB-silenced SB3×6 (left) and control Ctrl cultures (right); scale bar: 1 µm. b Cell size distribution of Ctrl, SB3×6, and SB2 cultures (OD₆₀₀ = 0.1). Bars in the graph represent the samples in the following order from left to right: Control (dark gray); SB3×6 (petrol); SB2 (orange). Cell sizes were measured using MicrobeJ software on phase contrast micrographs (n > 300, different biol. replicates). c FSC-histogram (Forward scatter, flow cytometry, created with Flowing Software 2.5.1) of silenced and control cultures sampled at OD₆₀₀ = 0.1 (n = 10,000 per biol. replicate). Bar chart represents percentage of cells in population exhibiting “small”, “average” and “large” size (size sections defined according to Ctrl histograms). Bars in the graph represent the samples in the following order from left to right (in every section): Control (dark gray); control-CC (light gray); SB3×2 (green); SB3×6 (petrol); SB3×6-CC (light petrol); SB2 (orange); SB2-CC (yellow). Error bars, mean ± SD (three biol. replicates). Asterisks indicate significant differences to Ctrl (two-tailed t test, n = 3, p ≤ 0.009) d Salt-stress experiment. Violin plot depicts decrease and increase of abundances of specific cell sizes of untreated (M) pIZ-plasmid SB3×6 and Ctrl cultures (n = 50 cells per replicate and condition), and upon 100 mm and 200 mm salt treatment (treatment upon salt addition = 45 min, 77 °C, agitating), respectively. Plots represent the samples in the following order from left to right (in each treatment): Control (gray); SB3×6 (petrol). Cell size differences were measured as in b). Source data are provided⁷⁰