FRAP analysis of RLK-GFP in pollen tubes expressing CA-rop1 or DN-rop1. (A and C) RLK-GFP was coexpressed with DN-rop1 (A) or CA-rop1 (C) in tobacco pollen tubes. After a 4-h incubation, transformed pollen tubes underwent FRAP analysis. The series of images shows before bleaching, immediately after bleaching (0 s), and recovery of fluorescence after bleaching at the indicated time points. The bleached area is marked by the boxes. Bars, 10 μm. (B) Quantitative FRAP analysis of A and C. The mean intensity of recovered fluorescence was measured by drawing a line on the photobleached PM regions and plotted as a function of time. Note the slow and weak fluorescence recovery in the DN-rop1–expressing tube. CA-rop1, FRAP of tube expressing CA-rop1. Control, FRAP of wild-type control tubes. (D) Quantitative FRAP analysis. The intensity of the recovered GFP signal at four PM regions (indicated by the numbers indicated in C) is plotted as a function of time. Note that fluorescence reappeared simultaneously at the whole bleached membrane. Recovered GFP signal was measured every 5 s for 80 s.