Fig. 2

A Diffusion of RibDG-mV with respect to existing subpopulations and subcellular localization of free and confined trajectories. A Jump distance (JD) analysis of RibDG-mV and RibDG-mV (ΔribH) SPT data using a two-component Rayleigh distribution model or a one-component Rayleigh distribution model (in gray). A non-linear least-squares fitting method has been applied to estimate their parameters, being the optimal number of components determined using statistical F-test for nested models. SPT data taken for this analysis and its results are summarized in Tables 1 and 2. B Model prediction-versus-observation plots for the Rayleigh distribution models of JDs in A. Data modeled based on Brownian motion are shown as a straight, red dotted line, the deviation of the observed data (fitted by double or single distributions, as indicated) to the modeled data is shown by the blue curved line, indicating very good agreement C Bubble plots displaying relative proportions of subpopulations for RibDG-mV and their according DCs that are derived from JD analysis in A. D Method for subcellular analysis of confined and free diffusing trajectories. (I) In the first step, all trajectories from a particular SPT dataset with at least five steps of consecutive localizations are projected onto a normalized 2D representation of the bacterial cytoplasm and transformed into a heat map displaying the overall spatial distribution of FP enzymes. (II) In the second step, the localization precision of each trajectory is estimated, an average value is calculated and further multiplied by a factor of 2.5 to yield the radius for a circle used for the analysis of confinement. (III) As a result of the analysis, two classes of trajectories arise (IV) which can be further transformed into separate heat maps. Trajectories can furthermore be analyzed in terms of transitions to confinement with respect to different numbers of confined steps yielding characteristic dwell times. E Subcellular analysis of confined and free trajectories shown for different numbers of consecutive steps projected onto the cytoplasm of a normalized cell representation. Displayed are the results using the radii given in Table S2 with three, six or nine consecutive steps of confinement