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[Preprint]. 2024 Dec 2:2024.12.02.626492. [Version 1] doi: 10.1101/2024.12.02.626492

Figure 6: SPEF1 stabilizes microtubules by binding to the seam.

Figure 6:

(A) Denoised (SIRT-like filter) tomogram of purified tip CP (native cilium data from Legal et al., 2023) and microtubules incubated with HsSPEF1-CH. Arrowheads point to a repetitive density found between both microtubules. Scale bar: 40 nm. (B) Subtomogram averaging map of the 13-PF and 14-PF microtubules incubated with HsSPEF1-CH overlapping with models of 13-PF and 14-PF microtubules binding to HsSPEF1-CH. The density corresponding to HsSPEF1-CH is coloured in green. (C) Representative kymographs of 750 nM recombinant HsSPEF1-CH and 8 μM MT. Horizontal scale bar: 4 μm; vertical scale bar: 5 minutes. (D - G) Microtubule length, lifetime, growth rate and rescue probability versus HsSPEF1-CH concentration. Each point represents the average, and the error bars represent the standard deviation. n=421, 482, 568, 643, 580, and 555 microtubules for HsSPEF1-CH concentrations of 0, 100, 400, 750, 1000, and 2000, respectively, obtained from three independent experiments.