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. 2023 Mar 9;6(5):e202201714. doi: 10.26508/lsa.202201714

Figure 1. D2 region in CAMSAP3 prefers GMPCPP-stabilized microtubules.

Figure 1.

(A) Schematic of human CAMSAP3 at its C-terminal. (B) Schematic of TIRF microscope used in the binding assay. The depolymerization of GDP-MTs was inhibited by adding 25% glycerol in the imaging buffer. We avoided using taxol in this assay because of its effects on the microtubule lattice. Blue indicates the GMPCPP-stabilized microtubule seed. Magenta shows GDP-MT elongated from the seed. (C) Representative images showing D2 binding to microtubules. Contrasts are set to the same range for each channel. Images after unmixing the fluorescence intensities are shown. Scale bar, 5 μm. 75 GMPCPP-MTs and 75 GDP-MTs from three replicates were analyzed for each D2 concentration. (D) Quantification of the average fluorescence intensity of D2-mCherry along microtubules. Colors represent three replicates. Error bars, S.D. ****P < 0.0001 (Welch’s t test).