Figure 5.
Dynein and its machinery travel to the distal tip of the axon separately. (A) Schematic of microfluidic device showing treatment in the somatodendritic compartment with JFX 554/650 ligand and imaging in the axonal compartment. Example kymographs of retrograde dynein (Halo-DYNC1H1), dynactin (Halo-ACTR10), LIS1 (PAFAH1B1-Halo), and NDEL1 (Halo-NDEL1) movement in 21–23 DPI neurons. (B) The frequency of anterograde motile events in dynein, dynactin, LIS1, and NDEL1 in 21–23 DPI neurons (dynein: 198 tracks, 6 videos, N = 5; dynactin: 211 tracks, 7 videos, N = 5; LIS1: 68 tracks, 5 videos, N = 5; NDEL1: 86 tracks, 15 videos, N = 3). Dynein versus LIS1: *P = 0.033, dynactin versus LIS1: **P = 0.0077, dynein versus dynactin: P = 0.60, dynactin versus NDEL1: P = 0.17, dynein versus NDEL1: P = 0.36, LIS1 versus NDEL1: P = 0.36, Kruskal–Wallis test, Dunn post hoc test. (C) The speed of anterograde dynein, dynactin, LIS1 and NDEL1 particles in 21–23 DPI neurons (dynein: 198 tracks, 6 videos, N = 5; dynactin: 211 tracks, 7 videos, N = 5; LIS1: 68 tracks, 5 videos, N = 5; NDEL1: 86 tracks, 15 videos, N = 3). Dynein versus dynactin: *P = 0.044; dynein versus NDEL1: **P = 0.0056; dynactin versus LIS1: *P = 0.015; LIS1 versus NDEL1: **P = 0.0018, dynein versus LIS1: P = 0.68, dynactin versus NDEL1: P = 0.30, Kruskal–Wallis test, Dunn post hoc test. (D) Example kymographs of anterograde dynein (Halo-DYNC1H1) and dynactin (DCTN4-SNAP) movement in 21–23 DPI neurons. Arrows point to colocalized dynein and dynactin. (E) The average number of tracks per experiment in 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). Boxplot shows median, first, and third quartiles. Upper/lower whiskers extend to 1.5× the interquartile range.

Dynein and its machinery travel to the distal tip of the axon separately. (A) Schematic of microfluidic device showing treatment in the somatodendritic compartment with JFX 554/650 ligand and imaging in the axonal compartment. Example kymographs of retrograde dynein (Halo-DYNC1H1), dynactin (Halo-ACTR10), LIS1 (PAFAH1B1-Halo), and NDEL1 (Halo-NDEL1) movement in 21–23 DPI neurons. (B) The frequency of anterograde motile events in dynein, dynactin, LIS1, and NDEL1 in 21–23 DPI neurons (dynein: 198 tracks, 6 videos, N = 5; dynactin: 211 tracks, 7 videos, N = 5; LIS1: 68 tracks, 5 videos, N = 5; NDEL1: 86 tracks, 15 videos, N = 3). Dynein versus LIS1: *P = 0.033, dynactin versus LIS1: **P = 0.0077, dynein versus dynactin: P = 0.60, dynactin versus NDEL1: P = 0.17, dynein versus NDEL1: P = 0.36, LIS1 versus NDEL1: P = 0.36, Kruskal–Wallis test, Dunn post hoc test. (C) The speed of anterograde dynein, dynactin, LIS1 and NDEL1 particles in 21–23 DPI neurons (dynein: 198 tracks, 6 videos, N = 5; dynactin: 211 tracks, 7 videos, N = 5; LIS1: 68 tracks, 5 videos, N = 5; NDEL1: 86 tracks, 15 videos, N = 3). Dynein versus dynactin: *P = 0.044; dynein versus NDEL1: **P = 0.0056; dynactin versus LIS1: *P = 0.015; LIS1 versus NDEL1: **P = 0.0018, dynein versus LIS1: P = 0.68, dynactin versus NDEL1: P = 0.30, Kruskal–Wallis test, Dunn post hoc test. (D) Example kymographs of anterograde dynein (Halo-DYNC1H1) and dynactin (DCTN4-SNAP) movement in 21–23 DPI neurons. Arrows point to colocalized dynein and dynactin. (E) The average number of tracks per experiment in 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). Boxplot shows median, first, and third quartiles. Upper/lower whiskers extend to 1.5× the interquartile range.

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