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. 2019 Oct 9;116(43):21789–21799. doi: 10.1073/pnas.1907397116

Fig. 6.

Fig. 6.

Kinetics of SpoIVA recruitment onto SpoVM-saturated SSLBs. (A) Schematic of an SSLB (gray) saturated with unlabeled SpoVM (blue) incubated with limiting amounts of Cy3-SpoIVA (magenta). (B) Adsorption of increasing concentrations of Cy3-SpoIVA onto SpoVM-saturated 2-µm SSLBs in vitro. Fluorescence intensity on each SSLB measured using flow cytometry. Data fit is as in Fig. 1. Data points are mean values; error bars: SD (n = ∼20,000). (C) SpoVM-saturated 2-μm SSLBs incubated with Cy3-SpoIVA visualized using bright field (Top); maximum-intensity projection of aggregate Cy3-SpoIVA molecule binding events over time (Middle); localization plot of total Cy3-SpoIVA localization events (green) or those that occur on SSLBs (blue) (Bottom). (D) Kymograph of Cy3-SpoIVA molecules binding on a SpoVM-saturated 2-μm SSLB observed for 200 frames. Short-lived (orange) and long-lived (blue) interactions are indicated. (E) Cumulative distribution function of SpoIVA duration on SpoVM-saturated SSLBs fit with 1-component (Left) or 2-component (Right) exponential decay model. Arrow: parameter space of data that was better fit with using a 2-component model. (F) Residence times of long-lived Cy3-SpoIVA bound to SpoVM-saturated 2-μm SSLBs at Cy3-SpoIVA concentrations indicated. Data points are one region of interest containing 3 to 14 SSLBs. (Scale bars: 2 μm.)