Figure 6. Monomeric N terminus of EB3 tracks growing MT ends in vitro. (A–D) In vitro MT plus-end tracking assay. Representative TIRFM images (A and C) and kymographs (B and D) show specific accumulation of GFP-EB3 (100 nM) and EB3-NL-mVenus (100 nM) at the growing, but not shortening MT plus (+) and minus (−) ends. Horizontal bars, 5 µm; vertical bar, 60 s. (E) Dual-color imaging of in vitro plus-end tracking assays performed with the indicated concentrations of EB3-NL-mVenus and mCherry-EB3. Images were processed by applying Blur filter in Photoshop. Bar, 1 µm. (F) Ratio of fluorescence intensity at the growing MT tip and on the MT lattice for the indicated protein mixtures (after background subtraction). Green plots show measurements for EB3-NL-mVenus and red plots for mCherry-EB3. 10–20 MT tips were measured per experiment; error bars represent SD.
Figure 6.

Monomeric N terminus of EB3 tracks growing MT ends in vitro. (A–D) In vitro MT plus-end tracking assay. Representative TIRFM images (A and C) and kymographs (B and D) show specific accumulation of GFP-EB3 (100 nM) and EB3-NL-mVenus (100 nM) at the growing, but not shortening MT plus (+) and minus (−) ends. Horizontal bars, 5 µm; vertical bar, 60 s. (E) Dual-color imaging of in vitro plus-end tracking assays performed with the indicated concentrations of EB3-NL-mVenus and mCherry-EB3. Images were processed by applying Blur filter in Photoshop. Bar, 1 µm. (F) Ratio of fluorescence intensity at the growing MT tip and on the MT lattice for the indicated protein mixtures (after background subtraction). Green plots show measurements for EB3-NL-mVenus and red plots for mCherry-EB3. 10–20 MT tips were measured per experiment; error bars represent SD.

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