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. 2021 Dec 31;10:e67282. doi: 10.7554/eLife.67282

Figure 5. SSNA1 detects microtubule lattice damage.

(A) Representative images of GMPCPP-stabilized microtubules (red) that were pre-incubated for 5 min with 100 nM spastin and 1 mM ATP and then subsequently incubated with 5 µM 647-SSNA1 (cyan). The images shown are from 8 min after SSNA1 addition. The orange asterisks indicate the microtubule used for kymograph and linescan analysis. (B) Kymographs showing SSNA1 localization to sites of spastin-induced microtubule damage. (C) Corresponding linescan analysis of the microtubule and SSNA1 intensities every minute from 0 min to 10 min after the introduction of SSNA1.

Figure 5—source data 1. An Excel sheet containing numerical data for the quantification of the SSNA1 and seed fluorescence intensity linescans as presented in Figure 5 and Figure 5—figure supplement 1.

Figure 5.

Figure 5—figure supplement 1. SSNA1 detects sites of microtubule lattice damage on GMPCPP-stabilized and Taxol-stabilized microtubules.

Figure 5—figure supplement 1.

(A) An image of a GMPCPP-stabilized microtubule that was incubated with 2 µM 488-SSNA1 and corresponding fluorescence intensity linescan. (B) An image of a Taxol-stabilized microtubule that was incubated with 5 µM 488-SSNA1 and corresponding fluorescence intensity linescan.
Figure 5—figure supplement 2. Spastin protein purification.

Figure 5—figure supplement 2.

SDS-Page gel of purified 6His-MBP-spastin and His-strep-sfGFP-spastin (del227) proteins used in this study.