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Figure S1. Morphological and growth analysis of wildtype BY-2 cells versus YFP-2xFYVE-transformed BY-2 cells. Of each cell line their increase in cell volume was followed for 7 days after transferring an equal cell volume and the breadth/length ratio was determined. Each day 5 ml was taken and cell volume was determined after centrifugation for 5 minutes at 107 g (experiments were carried out in triplicates). (a) DIC image of wild- type cells, (b) DIC image of YFP-2xFYVE cells, (c) growth curve of wild-type cells (solid line) versus YFP-2xFYVE cells (dashed line)and (d) breadth/length ratio of wild-type and YFP-2xFYVE cells (n=80 cells). Bars = 10 μm.

Movie S1. YFP-2xFYVE dynamics in cowpea protoplasts. A time-lapse animation taken from cowpea protoplasts transiently expressing YFP-2xFYVE allowing visualization of YFP-2xFYVE dynamics. Chlorophyll is in red, YFP-2xFYVE is in green, DIC is in gray. Time (sec) is shown in the movie.

Movie S2. YFP-2xFYVE dynamics in tobacco BY-2 cells. A time-lapse animation taken from YFP-2xFYVE expressing tobacco BY-2 cells allowing visualization of PtdIns3P dynamics. YFP-2xFYVE is in green, DIC is in gray. Total time (sec) is shown in the movie.

Movie S3. Transient interactions between YFP-2xFYVE labeled vesicles. A time-lapse animation taken from YFP-2xFYVE expressing tobacco BY-2 cells, suggesting that YFP-2xFYVE-labelled vesicles undergo fusion and fission. Shown as an inverted gray scale images. Time is shown (sec) in the movie.

Movie S4 and S5. YFP2xFYVE dynamics depend on the actin cytoskeleton. Time-lapse animations of YFP-2xFYVE expressing tobacco BY-2 cells, after addition of 1 μM Latrunculin A (S4) or oryzalin (10 μM) (S5). YFP-2xFYVE is in green and DIC is in gray. Total time is indicated in the movie.

Movie S6. Effect of PI-3 kinase inhibitor wortmannin on YFP-2xFYVE distribution. A time-lapse animation taken from YFP-2xFYVE expressing tobacco BY-2 cells directly after the addition of 10 μM wortmannin. YFP-2xFYVE is in green and DIC is in gray. Total time (sec) is indicated in the movie.

Movie S7. YFP-2xFYVE fluorescence partially co-localizes with the endocytic tracer FM4-64. A time-lapse animation taken from YFP-2xFYVE expressing tobacco BY-2 cells after a pulse labelling with FM4-64 (10 min, 4 μM). YFP-2xFYVE is in green, FM4-64 is in red. Total is indicated in the movie.

Movie S8. YFP-2xFYVE labels endocytic/prevacuolar vesicles. A time-lapse animation taken from tobacco BY-2 cells co-expressing YFP-2xFYVE and mRFP-AtRABF2b, showing a strong overlap of fluorescent signal. YFP-2xFYVE is in green, mRFP-AtRABF2b is in red. Total time in seconds is shown in movie.

Movie S9. YFP-2xFYVE does not co-localize with the Golgi marker STtmd-CFP. A time-lapse animation taken from tobacco BY-2 cells co-expressing YFP-2xFYVE and STtmd-CFP showing no overlap of fluorescent signal, although YFP-2xFYVE labeled vesicles were regular observed in close vicinity of STtmd-CFP labelled Golgi stacks (see Figure 6) YFP-2xFYVE is in green, STtmd-CFP is in red. Total time, in seconds, is shown in the movie.

Movie S10. YFP-2xFYVE dynamics in growing root hairs of Arabidopsis. A time-lapse animation taken from a growing root hair of a one week old Arabidopsis plant stably expressing YFP-2xFYVE. YFP-2xFYVE is in green, DIC is in gray. Total time (sec) is indicated in the movie.

Movie S11. YFP-2xFYVE dynamics in leaf epidermal cells of Arabidopsis. A time-lapse animation taken from leaf epidermal cells of an Arabidopsis plant expressing YFP-2xFYVE. YFP-2xFYVE is in green, DIC is in gray. Total time in seconds is shown in movie.

Movie S12. Image-stack taken from leaf epidermal cells of an YFP-2xFYVE expressing Arabidopsis plant. Total image stack compromises 19.5 μm.

Movie S13. YFP-2xFYVE dynamics in Arabidopsis guard cells. A time-lapse animation taken from a guard cell of an Arabidopsis plant expressing YFP-2xFYVE. YFP-2xFYVE is in green, DIC is in gray. Total time (sec) is shown in the movie.

Movie S14. Image-stack taken from a guard cell of an Arabidopsis plant expressing YFP-2xFYVE. YFP-2xFYVE is in green, DIC is in grey. Total image stack compromises 12 μm.

Movie S15. YFP-2xFYVE fluorescence during cytokinesis in tobacco BY-2 cells. A time-lapse animation taken from tobacco BY-2 cells expressing YFP-2xFYVE undergoing cell division. YFP-2xFYVE is in green, DIC is in grey. Total time in seconds is shown in movie.

Movie S16. Image stack of a dividing tobacco BY-2 cell expressing YFP-2xFYVE. An image-stack taken from a dividing tobacco BY-2 cell expressing YFP-2xFYVE. YFP-2xFYVE is in green, DIC is in grey. Total image stack compromises 23 μm.

Movie S17. YFP-2xFYVE fluorescence is absent from the cell plate. A time-lapse animation taken from a dividing tobacco BY-2 cells expressing YFP-2xFYVE in the presence of FM4-64. YFP-2xFYVE fluorescence follows, but does not label the forming cell plate. YFP-2xFYVE is in green, FM4-64 is in red. Total time of the movie is indicated (sec).

Movie S18. 4D-imaging of dividing tobacco BY-2 cells expressing YFP-2xFYVE. A 4D time-lapse animation taken from tobacco BY-2 cells expressing YFP-2xFYVE undergoing cell division in the presence of FM4-64 is shown. YFP-2xFYVE is in green, FM4-64 is in red. Total time (sec) is indicated in the movie.

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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.