Figure 3.

Polyglutamylation activates a short version of spastin in vivo. (A) HeLa cells transfected with the N-terminally truncated form of spastin, C389-spastin-EYFP (mutated D552N; Fig. S3), were grown for 15 h and subsequently transfected with either active TTLL11 or inactive mutant (TTLL11d) 7 h before fixation and immunofluorescence staining by 12G10. CFP coexpressed from the TTLL11 plasmids allowed identification of transfected cells. Bar, 20 µm. (B) Fraction of cells expressing C389-spastin D552N-EYFP and CFP showing MT severing (all severing phenotypes as shown in A were compiled). A minimum of 100 cells was analyzed in three independent experiments. Error bars SD. Significance was determined by two-tailed Student’s t test. All P values (**) are below 0.001.