Figure 6.

RFO1-GFP particle dwell time at the PM is altered by the perturbation of cell wall pectin methylation.

(A) Representative spinning-disc confocal images (top) of a 5-day-old root elongating epidermal cell and kymographs (bottom) along the length of the dashed yellow lines from the corresponding video (n = 50 frames, 1 frame/s) of RFO1-GFP (rfo1-1 pRFO1RFO1-GFP) and RFO1-GFP iPMEIox (rfo1-1 pRFO1RFO1-GFP iPMEIox) seedlings treated with 12 μM estradiol for 2 h. RFO1-GFP iPMEIox seedlings were co-treated with Mock (−) or 10 μg/ml of dmPectin (+). Unless specified, all data derived from 180-s videos taken at 1 frame/s. Scale bar, 5 μm.

(B) Dwell time frequencies of RFO1-GFP particles at the PM as depicted in (A). Means (μ) are shown by dashed lines. For each condition, data represent N > 12 000 particles from a minimum of three cells per root and three roots.

(C) Average RFO1-GFP particle dwell times at the PM.

(D) Quantification of the total number of RFO1-GFP particles at the PM (left), and particles with dwell times >50 s (middle) and >100 s (right).

(E) Average instantaneous diffusion coefficients of RFO1-GFP particles at the PM. (C–E) Data represent the mean ± SE of N > 25 cells per condition from three independent replicates using three cells per root, three roots per condition, and tracking ≥1072 particles/cell, obtained from videos as described in (A). One-way ANOVA with Dunnett’s multiple-comparison test, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.