Live-cell single-molecule analysis of autophagy proteins. (A and B) Example of single-particle tracking of (A) ATG9 and (B) ATG16, and the corresponding fitting of the step-size probability distribution with SpotON algorithm. Numbers inside the micrographs indicate the imaging frame associated with the track. Movies were acquired at 6.8 ms per frame, scale bar = 1 μm. (C) Results of diffusive analysis for the HaloTagged autophagy proteins under control and EBSS starvation. Top three panels present the diffusion coefficients of the tracks based on the SpotON analysis. Bottom panel depicts the percentage associated with each fraction. The box indicates confidence interval ± SD, the square indicates the average, and the horizontal line is the median; for each condition, three biological replicates were analyzed, ∼20 cells/replicate.
Figure 3.

Live-cell single-molecule analysis of autophagy proteins. (A and B) Example of single-particle tracking of (A) ATG9 and (B) ATG16, and the corresponding fitting of the step-size probability distribution with SpotON algorithm. Numbers inside the micrographs indicate the imaging frame associated with the track. Movies were acquired at 6.8 ms per frame, scale bar = 1 μm. (C) Results of diffusive analysis for the HaloTagged autophagy proteins under control and EBSS starvation. Top three panels present the diffusion coefficients of the tracks based on the SpotON analysis. Bottom panel depicts the percentage associated with each fraction. The box indicates confidence interval ± SD, the square indicates the average, and the horizontal line is the median; for each condition, three biological replicates were analyzed, ∼20 cells/replicate.

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