Cellulose synthase-like D movement in the plasma membrane requires enzymatic activity

mEGFP-CesA10 and mEGFP-CSLD6 particles were imaged with VAEM every 2s for 10-20 minutes. Images were processed before tracking using subtract background and enhance contrast in Fiji. Particle tracking was performed with the Fiji plugin TrackMate (Tinevez et al., 2017). For CesA10, stacks were reduced by 10, resulting in 20 s time intervals between frames. For CSLD6, stacks were reduced by 6, resulting in 12 s time intervals between frames. LoG detector (estimate blob diameter 0.3 µm) and LAP tracker (frame to frame linking max distance 0.5 µm; Penalties: Quality=1; no gap closing allowed) were selected. The filters on tracks were set as follows: duration >60 s; track displacement >0.5 µm. Mean speed and the confinement ratio of tracks were obtained and plotted in Figure 3. The confinement ratio is a ratio of the straight-line distance between the start and end points of the trajectory divided by the actual trajectory distance. The closer the confinement ratio value is to one, the more linear the trajectory.