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. 2018 Jun 26;7:e36306. doi: 10.7554/eLife.36306

Figure 8. Comparison of motile mRNPS reconstituted with WT dynein or dynein without LC8.

Figure 8.

(A) SDS-PAGE of (lane 1) molecular mass markers, (lane 2) expressed WT dynein, (lane 3) dynein expressed without LC8. The identity of the bands labeled DIC and DLIC was confirmed by mass spectrometry. The band marked with an asterisk is a FLAG-reactive fragment that was shown by mass spectrometry to be derived from the heavy chain. DHC, dynein heavy chain; DIC, dynein intermediate chain; DLIC, dynein light intermediate chain. Higher loads of the same samples are shown in the inset so that the light chains can be visualized. 4–12% SDS-PAGE, MES buffer. (B) WT dynein speeds (filled red circles) (0.42 ± 0.13 μm/s and 0.71 ± 0.34 μm/s, n = 396) were not statistically different from dynein minus the LC8 light chain (open blue squares) (0.40 ± 0.13 μm/s and 0.79 ± 0.39 μm/s, n = 488; p=0.57, t-test, mean ± SD). (C) WT dynein run lengths (6.2 ± 0.11 μm, n = 113) were the same as dynein without LC8 (6.9 ± 0.14 μm, n = 102 r; p=0.46, Kolmogorov–Smirnov test, mean ± SE). Complexes were reconstituted with two dyneins per dynactin. See Figure 8—source data 1.

Figure 8—source data 1. Dataset for Figure 8.
elife-36306-fig8-data1.xlsx (11.3KB, xlsx)
DOI: 10.7554/eLife.36306.026