G1/S single particle trajectories length and jump size distributions. (A and B) Trajectory length distributions for sptPALM on (A) Swi4-mEos3.2 and (B) Mbp1-mEos3.2 in live cells. Data are shown for Swi4-mEos3.2 and Mbp1-mEos3.2 in cells grown in SC + 2% glucose. Results were identical for cells grown in SC + 2% glycerol. JDDs for Swi4 (dotted lines), Mbp1 (filled lines), and Swi6 (dashed lines) single molecules in fixed cells (C and E) as a control for instrument jitter and (D and F) in live cells. Colors in all JDD distributions correspond to cells grown in SC + 2% glucose medium (blue) and SC + 2% glycerol medium (red). (C and D) Distributions of jump distances over large times (570 ms and 510 ms, respectively, for fixed and live cells with 20 and 18 points, respectively). (E and F) distributions of jump distances over shorter times (210 ms, eight-point trajectories for both fixed and live cells). Red arrows indicate occurrences of large jumps in live cells distributions that exceed confined diffusion (black arrows).