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. 2018 Sep 26;9:3942. doi: 10.1038/s41467-018-06310-1

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© The Author(s) 2018

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. 4 — MinDE induce oscillatory and time-averaged concentration gradients of model membrane proteins in microcompartments. a Experimental setup: PDMS-microcompartments are lined with an SLB and covered by air to confine the proteins. b Representative time-lapse images and kymographs of MinDE oscillations and streptavidin counter-oscillations in the compartments (1 µM MinD, 2 µM MinE, streptavidin-Alexa647). Brightness of the streptavidin channel was corrected for bleaching using histogram matching in Fiji. Scale bars: 10 µm. c Time-averaged fluorescence intensity profiles of MinDE (green) and streptavidin (cyan) oscillation in b showing clear concentration gradients for both MinD and streptavidin. d Time-averaged fluorescence intensity profiles (gray lines) for EGFP-MinD and streptavidin aligned and projected to a unit box (see Supplementary Fig. 14 for details). Bold, colored lines represent the mean profiles, generated from three independent experiments with N = 35 microcompartments. e Representative time-lapse images and kymographs of MinDE oscillations and mCh-MTS(BsD) counter-oscillations in PDMS microcompartments (1 µM MinD (30% EGFP-MinD), 2 µM MinE, 0.5 µM mCh-MTS(BsD)). Scale bars: 10 µm. f Time-averaged fluorescence intensity profiles of MinDE (green) and mCh-MTS(BsD) (magenta) oscillations in e showing a clear protein gradient for MinD and homogenous protein distribution of mCh-MTS(BsD). g Time-averaged fluorescence intensity profiles (gray lines) for EGFP-MinD and mCh-MTS(BsD) aligned and projected to a unit box. Bold, colored lines represent the mean profiles, generated from three independent experiments with in total N = 45 microcompartments. h Schematic explaining how the MinDE system positions lipid-anchored streptavidin and mCh-MTS constructs in rod-shaped microcompartments. MinDE oscillations drive counter-oscillations of lipid-anchored streptavidin and mCh-MTS constructs, thereby establishing a time-averaged concentration gradient of lipid-anchored streptavidin with maximal concentration in the geometric center, but no concentration gradient of mCh-MTS