Figure S2.
Analysis of the dynein machinery in the axon. (A) The amount of time until the first processive retrograde fluorescent particle was detected in dynein, dynactin, LIS1, and NDEL1 21–23 DPI neurons (dynein: 8 videos, N = 8; dynactin: 6 videos, N = 6; LIS1: 5 videos, N = 5; NDEL1: 2 videos, N = 2). Dynein versus dynactin: P = 0.67, NDEL1 versus dynactin: P = 0.53, LIS1 versus dynactin: P = 0.72, dynein versus NDEL1: P = 0.72, dynein versus LIS1: P = 0.99, NDEL1 versus LIS1: P = 0.73. (B) The amount of time dynein, dynactin, LIS1, and NDEL1 spent pausing in the retrograde direction (dynein: 162 tracks, 21 videos, N = 6; dynactin: 68 tracks, 10 videos, N = 4; LIS1: 38 tracks, 14 videos, N = 6; NDEL1: 24 tracks, 10 videos, N = 3). Dynein versus dynactin: P = 0.18, NDEL1 versus dynactin: P = 0.42, LIS1 versus dynactin: P = 0.09, dynein versus NDEL1: P = 0.71, dynein versus LIS1: P = 0.68, NDEL1 versus LIS1: P = 0.48. Pauses are defined as spots moving slower than 0.1 µm/s. (C) The average length of time each pause lasted for dynein, dynactin, LIS1, and NDEL1. Dynein versus dynactin: P = 0.3, NDEL1 versus dynactin: P = 0.63, LIS1 versus dynactin: P = 0.50, dynein versus NDEL1: P = 0.68, dynein versus LIS1: P = 0.06, NDEL1 versus LIS1: P = 0.26. Kruskal–Wallis test, Dunn post hoc test. (D) Instantaneous retrograde velocities of dynein, dynactin, LIS1, and NDEL1 particles in 21–23 DPI neurons. (E) The percent of time dynein, dynactin, LIS1, and NDEL1 spent pausing. Dynein versus dynactin: *P = 0.044; dynein versus NDEL1: *P = 0.026; dynactin versus LIS1: **P = 0.0037; LIS1 versus NDEL1: **P = 0.0028, dynactin versus NDEL1: P = 0.63, dynein versus LIS1: 0.37. Kruskal–Wallis test, Dunn post hoc test. Pauses are defined as spots moving slower than 0.1 µm/s. (F) Average length of time each pause lasted for dynein, dynactin, LIS1, and NDEL1 in the anterograde direction. Dynein versus dynactin: P = 0.15, NDEL1 versus dynactin: P = 0.95, LIS1 versus dynactin: P = 0.09, dynein versus NDEL1: P = 0.18, dynein versus LIS1: P = 0.81, NDEL1 versus LIS1: P = 0.13. Kruskal–Wallis test, Dunn post hoc test. (G) Instantaneous anterograde velocities of dynein, dynactin, LIS1 and NDEL1 particles in 21–23 DPI neurons. (H) The average speed of anterograde dynein and dynactin particles in dual labelled 21–23 DPI neurons. (dynein: 413 tracks, 25 videos, N = 3; dynactin: 732 tracks, 25 videos, N = 3; Co-loc: 57 tracks, 25 videos, N = 3; dynein versus dynactin: ***P = 0.000056, dynein versus co-loc: *P = 0.042, dynactin versus co-loc: ***P = 0.00034. One-way ANOVA, Tukey’s post hoc test.) (I) The percent colocalization between dynein and dynactin tracks. Boxplots show median, first, and third quartiles. Upper/lower whiskers extend to 1.5× the interquartile range.

Analysis of the dynein machinery in the axon. (A) The amount of time until the first processive retrograde fluorescent particle was detected in dynein, dynactin, LIS1, and NDEL1 21–23 DPI neurons (dynein: 8 videos, N = 8; dynactin: 6 videos, N = 6; LIS1: 5 videos, N = 5; NDEL1: 2 videos, N = 2). Dynein versus dynactin: P = 0.67, NDEL1 versus dynactin: P = 0.53, LIS1 versus dynactin: P = 0.72, dynein versus NDEL1: P = 0.72, dynein versus LIS1: P = 0.99, NDEL1 versus LIS1: P = 0.73. (B) The amount of time dynein, dynactin, LIS1, and NDEL1 spent pausing in the retrograde direction (dynein: 162 tracks, 21 videos, N = 6; dynactin: 68 tracks, 10 videos, N = 4; LIS1: 38 tracks, 14 videos, N = 6; NDEL1: 24 tracks, 10 videos, N = 3). Dynein versus dynactin: P = 0.18, NDEL1 versus dynactin: P = 0.42, LIS1 versus dynactin: P = 0.09, dynein versus NDEL1: P = 0.71, dynein versus LIS1: P = 0.68, NDEL1 versus LIS1: P = 0.48. Pauses are defined as spots moving slower than 0.1 µm/s. (C) The average length of time each pause lasted for dynein, dynactin, LIS1, and NDEL1. Dynein versus dynactin: P = 0.3, NDEL1 versus dynactin: P = 0.63, LIS1 versus dynactin: P = 0.50, dynein versus NDEL1: P = 0.68, dynein versus LIS1: P = 0.06, NDEL1 versus LIS1: P = 0.26. Kruskal–Wallis test, Dunn post hoc test. (D) Instantaneous retrograde velocities of dynein, dynactin, LIS1, and NDEL1 particles in 21–23 DPI neurons. (E) The percent of time dynein, dynactin, LIS1, and NDEL1 spent pausing. Dynein versus dynactin: *P = 0.044; dynein versus NDEL1: *P = 0.026; dynactin versus LIS1: **P = 0.0037; LIS1 versus NDEL1: **P = 0.0028, dynactin versus NDEL1: P = 0.63, dynein versus LIS1: 0.37. Kruskal–Wallis test, Dunn post hoc test. Pauses are defined as spots moving slower than 0.1 µm/s. (F) Average length of time each pause lasted for dynein, dynactin, LIS1, and NDEL1 in the anterograde direction. Dynein versus dynactin: P = 0.15, NDEL1 versus dynactin: P = 0.95, LIS1 versus dynactin: P = 0.09, dynein versus NDEL1: P = 0.18, dynein versus LIS1: P = 0.81, NDEL1 versus LIS1: P = 0.13. Kruskal–Wallis test, Dunn post hoc test. (G) Instantaneous anterograde velocities of dynein, dynactin, LIS1 and NDEL1 particles in 21–23 DPI neurons. (H) The average speed of anterograde dynein and dynactin particles in dual labelled 21–23 DPI neurons. (dynein: 413 tracks, 25 videos, N = 3; dynactin: 732 tracks, 25 videos, N = 3; Co-loc: 57 tracks, 25 videos, N = 3; dynein versus dynactin: ***P = 0.000056, dynein versus co-loc: *P = 0.042, dynactin versus co-loc: ***P = 0.00034. One-way ANOVA, Tukey’s post hoc test.) (I) The percent colocalization between dynein and dynactin tracks. Boxplots show median, first, and third quartiles. Upper/lower whiskers extend to 1.5× the interquartile range.

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