Figure 8.

Moderate ectopic expression of mutated spastin isoforms does not affect MT repolymerization. After 15 min of depolymerization in the presence of 2 μg/ml nocodazole, cells were rinsed and cultured for 20 min in nocodazole-free medium to allow the repolymerization of MTs. The measurements of fluorescence intensity after staining with anti-tubulin antibody were used to evaluate MT levels. A, A full repolymerization of MTs was observed in control nonexpressing cells (outlined in yellow). B, MT levels in cells expressing moderate levels of M87 C448Y (outlined in red) were indistinguishable from MT levels in nonexpressing cells (outlined in yellow). C, Moderate levels of M1 C448Y colocalizing with MTs (cells outlined in red) did not affect MT repolymerization. D, Quantification of MT levels measured as tubulin fluorescence intensity shows no statistically significant differences between control cells and cells expressing mutated spastin isoforms. Scale bar, 20 μm. Values are mean ± SEM; n = 30.