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. 2012 Apr 3;10(4):e1001299. doi: 10.1371/journal.pbio.1001299

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Nagawa et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Time-lapse imaging of PIN1-GFP transiently expressed in tobacco PCs. PIN1-GFP signal was imaged using laser scanning confocal microscopy 3 d after the infiltration of tobacco leaves with agrobacterium containing pPIN1-PIN1-GFP. Internalization of PIN1-GFP from the PM (arrowheads) occurred preferentially in the indention region but not in the lobe region (arrow) where stronger PM accumulation of PIN1-GFP was observed. Note that only a cell at the left side of the image expressed PIN1-GFP, and internalization events were only visualized in that cell but not in the cell at the right side of the images. (B) A time-course analysis of PIN1-dendra2 internalization after photo-conversion in tobacco PCs. Signal in the red color represents photo-converted PIN1-dendra2. PIN1-dendra2 was transiently expressed in tobacco PCs for 3 d before photo-conversion was achieved. Coexpression of DN-ROP2 promoted vesicle formation (arrowheads) from the PM and accelerated decrease of PM signal. (C) Quantitative analysis of the PM signal representing photo-converted PIN1-dendra2 shown in (B). Relative intensity was measured as absolute value of intensity divided by value of intensity in the first time point. Error bars represent standard deviation (SD) (n = 5). (D) A time-course analysis of PIN1-dendra2 internalization after photo-conversion in Arabidopsis PCs. PIN1-dendra2 (and CA-ROP2) was transiently expressed in WT or rop4 R2i Arabidopsis PCs for 24 h. Formation of vesicles (arrowheads) and decrease of PM signal were accelerated in rop4 R2i cells. Coexpressing CA-ROP2 suppressed both vesicle formation and decrease of PM signal in rop4 R2i cells (n = 5). (E) Quantitative analysis of PM signal shown in (D). Measurements of signal were done as in (C).