Figure 5. Kymographs of early endosomes. Analysis of endosomal movement using kymographs: High-density-of-frame time-lapse sequences in the GFP channel were acquired over the time indicated in the vertical axes. Fluorescent signals detected across a line traced along the cytoplasm of hyphae, wide enough to cover the complete width of the cell, were plotted against time. In this type of representation, vertical lines correspond to static signals, whereas moving structures appear as diagonals, whose slope indicates the speed and direction of movement. (A) Kymograph showing early endosomes visualized with GFP-RabB. The prominent endosome indicated with asterisks moves in one direction for 17 μm before abruptly shifting movement toward the opposite direction. (B) As above, using GFP-RabA. In one of the two examples shown (*), an endosome moves for 16 μm before becoming essentially static for the remaining period of observation. Another endosome (**) enters the region analyzed, moving in the opposite direction, and does not slow down over the remaining duration of the time-series.