Figure 5. Kinesin-1 is regulated by MAP7 proteins in vitro. (A) Overview of full-length kinesin-1 (KIF5B) domains. (B) Streptavidin pull-down assay with extracts of HEK293T cells overexpressing BirA, the indicated KIF5B-GFP truncations (prey), and Bio-mCherry-MAP7 (bait) analyzed by Western blotting. (C) Kymographs of GFP-tagged full-length KIF5B (kinesin-1) on dynamic MTs in control conditions or in the presence of MAP7 or MAP7D3. (D) Quantification of kinesin-1 landing frequencies per MT and corrected for MT length, time of acquisition, and kinesin concentration. n = 99, 26, and 38 MTs from two or three independent experiments. (E) Histograms of run lengths fitted to a single exponential decay (red) with indicated rate constants (tau) as a measure of mean run length. n = 71, 542, and 568 kinesin runs from two or three independent experiments. (F) Gaussian fits of kinesin velocities. Histograms are shown in Fig. S3 G.
Figure 5.

Kinesin-1 is regulated by MAP7 proteins in vitro. (A) Overview of full-length kinesin-1 (KIF5B) domains. (B) Streptavidin pull-down assay with extracts of HEK293T cells overexpressing BirA, the indicated KIF5B-GFP truncations (prey), and Bio-mCherry-MAP7 (bait) analyzed by Western blotting. (C) Kymographs of GFP-tagged full-length KIF5B (kinesin-1) on dynamic MTs in control conditions or in the presence of MAP7 or MAP7D3. (D) Quantification of kinesin-1 landing frequencies per MT and corrected for MT length, time of acquisition, and kinesin concentration. n = 99, 26, and 38 MTs from two or three independent experiments. (E) Histograms of run lengths fitted to a single exponential decay (red) with indicated rate constants (tau) as a measure of mean run length. n = 71, 542, and 568 kinesin runs from two or three independent experiments. (F) Gaussian fits of kinesin velocities. Histograms are shown in Fig. S3 G.

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