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. 2018 Oct 18;14(10):e1007726. doi: 10.1371/journal.pgen.1007726

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© 2018 Rohs et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 6 — A. Histograms of velocity measurements determined for individual traces of ^SWMreB-mNeon (attλHC897) in wild-type (22±6 nm/s, n = 1467; black) and PBP2(L61R) cells [PR78] (25±7 nm/s, n = 949; turquoise). B. Histograms of velocity measurements determined for individual tracks of msfGFP-PBP2 (attλHC943) (18.9±8.1 nm/s, n = 2692; black) and msfGFP-PBP2(L61R) (attλPR128) (17.8±7.5 nm/s, n = 3440; turquoise) in ΔpbpA cells. Bin size, 5 nm/s. C. Kymographs of individual ^SWMreB-mNeon (attλHC897) tracks in wild-type (i) and PBP2(L61R) (ii) cells are displayed atop kymographs of individual msfGFP-PBP2 (attλHC943, i) or msfGFP-PBP2(L61R) (attλPR128, ii) tracks in ΔpbpA cells. Scale bar, 250nm. D. Violin plots for the number of ^SWMreB-mNeon tracks measured per cell area (um²) in wild-type (2.8±1.58, n = 609; black) and PBP2(L61R) cells (3.85±2.14, n = 300; turquoise) at 37°C. p<1e-10, Mann-Whitney U. The distribution of values along the x-axis capture the frequency of measurements along the y-axis. Lines designate quartiles with the dotted line indicating the mean value. Outliers are highlighted in red. E. Violin plots for the number of msfGFP-PBP2 (1.16±0.84, n = 581; black) and msfGFP-PBP2 (1.92±1.19, n = 517; turquoise) tracks measured per cell area (um²) in ΔpbpA cells at 30°C. As expected [36] there are less directionally moving rod complexes at lower temperatures. p<1e-29, Mann-Whitney U. F. Violin plots illustrating the distribution of normalized fluorescence measurements for ^SWMreB-mNeon (attλHC897) expressed in wild-type (0.83±0.36, n = 397) and PBP2(L61R) cells (0.85±0.31, n = 321). The fluorescence intensity acquired under TIRF illumination for individual cells was integrated and divided by similar measurements taken under widefield illumination, providing an approximation for the relative abundance of surface-associated ^SWMreB-mNeon. G. Representative SIM-TIRF micrographs of ^SWMreB-mNeon integrated at the native locus in wild-type [JAB593] and PBP2(L61R) cells [JAB576]. The signal for ^SWMreB-mNeon is pseudocolored green and overlaid a contrast-adjusted phase-contrast image. Scale bar, 1μm. H. Distributions of ^SWMreB-mNeon polymer lengths in wild-type [JAB593] (520±190nm, n = 502; black) and PBP2(L62R) cells [JAB576] (360±130nm, n = 614; turquoise). Bin size, 120nm. p<1e-53, Mann-Whitney U. I. Representative phase-contrast micrographs of wild-type [MG1655] or PBP2(L61R) [PR78] cells after a 4hr treatment with 2 μg/mL A22, an MreB-inhibitor. The minimum inhibitory concentration (MIC) of A22 for each cell type is displayed in μg/mL.