FIGURE 8:

SVs rigorously bound by KinA^G97E massively decorate MTs after latrunculin B treatment. (A) Localization of SVs labeled with GFP-RabE^RAB11 in the wt and in cells expressing rigor mutant kinesins with and without latB treatment. (B) Kymographs (10 µm long, starting at the apex) derived from time stacks of GFP-RabE^RAB11 images obtained from kinA^G97E and uncA^G116E cells treated with latB (600 frames/min). (C) Flux of anterograde and retrograde runs in regions described in B in five latB-treated hyphae for each genotype. The total number of runs per hypha and minute was determined by manual tracking on kymographs. Runs were classified as anterograde or retrograde and plotted as means with 95% CIs. The different data sets were compared by ANOVA followed by Tukey’s multiple comparison posttest. ***p < 0.001; n.s., nonsignificant. (D) KinA^G97E cells coexpressing mCh-RabA^RAB5 to label EEs and GFP-RabE^RAB11 to label SVs. In the untreated KinA^G97E control, SVs accumulate in the SPK and EEs in a slightly subapical aggregate. LatB treatment of KinA^G97E cells shifts SVs from the SPK to MTs, which are conspicuously decorated with rigorously bound vesicles. Note that the tip accumulation of EEs is not dispersed by actin depolymerization (see also Supplemental Movies S15 and S16).