Figure 3.

FRAP-based lateral diffusion measurements of cargos at apical, basal, outer, and inner domains. (a) Cartoon of the experimental set-up for estimating lateral diffusion indexes as defined by the evolution ratio of the average signal intensity of ‘x’ (nonbleached PM) over ‘y’ (bleached PM). Each region was 2 μm long. Relative fluorescence intensities from 0 (black) to 250 (bright/white) are represented by the color code. Scale bar=6 μm. (b, c) Evolution of signal ratio ‘x/y’ over 30 min for PIN1-GFP, PIN2-GFP, GFP-ABCG37, ABCG36-GFP, BOR1-GFP, and PIP2-GFP in control conditions (b) or under conditions where active processes were blocked (-e, 0.02% sodium azide and 50 mM 2-deoxy-D-glucose, and 50 μm cycloheximide; 45 min pretreatment) (c). The signal values of prebleach and postbleach fluorescence intensities were normalized and are s.e.m., n=4–5 FRAP experiments on different roots.