Figure 4.

MIT mutant forms of spastin show differential recruitment to VPS4-E235Q endosomes. GFP-VPS4-E235Q was transiently transfected into cell lines stably expressing myc-tagged wild-type spastin (A) or spastin mutants (B–F), and then cells were fixed and labeled with an anti-myc antibody. Insets show higher magnification views of the boxed areas. (G) Co-localization between GFP-VPS4-E235Q and spastin proteins was estimated by calculating the Pearson's correlation coefficient for red and green pixels in each cell, using Volocity software (n = 4 experiments, 15 cells per condition quantified in each experiment). Mean and SEM are shown, p-values vs. M87 wild-type spastin were calculated using one-way ANOVA with Dunnett's multiple comparison posttest. Scale bar = 10 μm.