Figure 1.

Sequential recruitment of kinesin and myosin-V in COS7 cells. (A) Assay: Sequential recruitment of kinesin and myosin-V by addition of gibberellin and rapalog, respectively. (B) Overview of constructs. MD, motor domain; CC, coiled coil; NC, neck coil; GAI, gibberellin insensitive; GID1, Gibberellin insensitive dwarf1. (C) Peroxisome distribution before recruitment of motors and after sequential recruitment of kinesin and myosin-V. Red curves indicate cell outline. Panels show individual frames of a cut out. Scale bar, 20 μm. (D) Radial kymograph indicating the redistribution of fluorescent peroxisomes relative to the cell axis. Vertical lines indicate addition of gibberellin (black) and rapalog (blue). Scale bar, 10 μm. (E) Displacement (expressed in R_90%) and correlation (frame-to-frame similarity from 0 to 1) vs. time for cells without added ligands (control, gray), cells with addition of gibberelin only (Gibberelin, black), and cells with addition of gibberellin and rapalog (Gib + rapa, red). N = 12, 16, and 14 cells for control, gibbellin, and gib+rapa groups, respectively. Data was obtained from 2 experiments, mean ± s.e.m. Vertical lines indicate time of addition of gibberellin (black) and rapalog (blue), if added. (F) Averages of ten frames of each cell at t = 0–4.5 min (before, B), t = 25–29.5 min (middle, M), and t = 55–59.5 min (end, E). ^*P < 0.05, ^***P < 0.001, ^****P < 0.0001, Friedman test, Dunn's post-hoc test.