Fig. 1.

MinDE can spatiotemporally regulate a model peripheral membrane protein. a mCh-MTS(BsD), mCherry fusion to the C-terminal amphipathic helix of B. subtilis MinD, homogenously covers SLBs in the absence of MinDE (1 µM mCh-MTS(BsD)). In the presence of MinDE and ATP mCh-MTS(BsD) forms traveling surface waves that are anticorrelated to the MinDE wave (1 µM mCh-MTS(BsD), 1 µM MinD (30% EGFP-MinD), 1 µM MinE). Scale bars: 50 µm. b Kymographs of the line selections shown in a. Scale bars: 50 µm and 100 s. c Intensity profiles of the line selections shown in a. mCh-MTS(BsD) fluorescence (magenta) on the SLBs in the presence of MinDE is reduced and shows clear maxima in the minima of the MinDE waves (min(MinD)) and clear minima in the MinDE wave maxima (max(MinD)). d Schematic of the analysis process. EGFP-MinD images are segmented to generate two binary masks that are subsequently multiplied with mCh-MTS(BsD) images to obtain average intensities for the full image and in the minimum and maximum of the MinDE wave. e Intensity ratio of the average fluorescence of mCh-MTS(BsD) in the presence over in the absence of MinDE. Intensity ratios are shown for the average intensity of the full image (${\mathit{I}}^{\mathrm{mCh}-\mathrm{MTS}\left(\mathrm{BsD}\right)}$), in the MinDE minimum (${\mathit{I}}_{\min \left(\mathrm{MinD}\right)}^{\mathrm{mCh-MTS(BsD)}}$) and in the MinDE maximum (${\mathit{I}}_{\max \left(\mathrm{MinD}\right)}^{\mathrm{mCh-MTS(BsD)}}$). Each data point (exp 1–3) is generated from at least one time series consisting of 75 images in one sample chamber. Cross and error bars depict the mean values and standard deviations from three independent experiments