Fig. 5: FRAP in concert with single-particle tracking informs on the material state of foci in bacterial cells.
From: An experimental framework to assess biomolecular condensates in bacteria
a Fluorescence recovery of mCherry-fusion proteins after photobleaching. Magenta circles indicate the FRAP region. Scale bar: 2 µm. b Quantification of fluorescence recovery of mCherry fusion proteins. Shading represents the standard error of the mean. cIagg: n = 14 foci; PopTagSL: n = 12 foci; PopTagLL: n = 14 foci; McdB: n = 10 foci. c Representative single-molecule trajectories. Overlay of tracks, color-coded according to the track apparent diffusion coefficient, obtained from representative E. coli cells expressing PAmCherry fusion proteins. Scale bars: 1 µm. d Diffusion coefficients of PAmCherry fusion proteins. The diffusion coefficients of the indicated PAmCherry fusions were obtained by fitting the histograms of the log diffusion coefficients of single tracks to a two-component Gaussian mixture model assuming that the fusion proteins are composed of a slow (closed circles) and fast (open circles) fraction. e Weight fractions of mobility states of PAmCherry fusion proteins. Two-component Gaussian mixture fitting results show an increase in slow mobility fraction (solid bars) and a decrease in fast mobility fraction (empty bars) as protein concentration increases. Source data are provided as a Source Data file.
