Figure 6. Cadherin-2 is transported by... | Rockefeller University Press

Figure 6.

$Cadherin-2 is transported by dynein and Kif5C in THNs.(A–C) When motors or vesicle formation is impaired, Cadherin-2 particles spend less time in motion. For full statistical testing, see Table S2. n = 682 tracks for drug control, n = 711 tracks for Brefeldin A, n = 287 tracks for Dynasore, n = 416 tracks for injection control, n = 308 tracks for p50-expressing embryos, n = 593 tracks for Kif5C-expressing embryos, n = 441 tracks Kif5C T94N-expressing embryos, n = 373 tracks for Rab11a-expressing embryos, and n = 152 tracks for Rab11a S25N-expressing embryos. Instantaneous velocities in the anterograde (B) as well as retrograde (C) direction are reduced when vesicle formation or motor transport is impaired. For full statistical testing, see Table S3 and Table S4. Anterograde: n = 2,728 values for drug control, n = 2,978 values for Brefeldin A, n = 1,451 values for Dynasore, n = 2,059 values for injection control, n = 1,044 values for p50-expressing embryos, n = 2,808 values for Kif5C-expressing embryos, n = 2,055 values Kif5C T94N-expressing embryos, n = 1,815 values for Rab11a-expressing embryos, and n = 837 values for Rab11a S25N-expressing embryos. Retrograde: n = 2,687 values for drug control, n = 3,034 values for Brefeldin A, n = 1,574 values for Dynasore, n = 2,127 values for injection control, n = 1,117 values for p50-expressing embryos, n = 2,791 values for Kif5C-expressing embryos, n = 2,132 values Kif5C T94N-expressing embryos, n = 1,740 values for Rab11a-expressing embryos, and n = 801 values for Rab11a S25N-expressing embryos. (D) Plotting maximal and mean velocities for each of the 416 particle tracks in control THNs illustrates that the majority of Cadherin-2 particles move only at low mean and maximal velocities, with some particles exhibiting higher mobility in both directions. (E) Converting the mean-maximal velocities plots into data density plots reveals that the fraction of slow-moving particles increases when vesicle formation is impaired by drugs. Color code is indicated on the right, where dark red indicates high data density. For comparison of the spread in the datasets, the area of the 0.8 density is outline in gray for each condition, and the area covered by this density level is indicated in the top left corner. The number of particle tracks for each condition is listed in the bottom left corner. Vertical gray lines separate areas without data content. (F) Similar results are obtained when vesicle formation or motor transport of Cadherin-2 particles are genetically targeted. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. All data shown in A–F represent n = 13 embryos for drug control, n = 19 embryos for Brefeldin A, n = 8 embryos for Dynasore, n = 13 embryos for injection control, n = 14 p50-expressing embryos, n = 11 Kif5C-expressing embryos, n = 16 Kif5C T94N-expressing embryos, n = 16 Rab11a-expressing embryos, and n = 7 Rab11a S25N-expressing embryos.$

Cadherin-2 is transported by dynein and Kif5C in THNs. (A–C) When motors or vesicle formation is impaired, Cadherin-2 particles spend less time in motion. For full statistical testing, see Table S2. n = 682 tracks for drug control, n = 711 tracks for Brefeldin A, n = 287 tracks for Dynasore, n = 416 tracks for injection control, n = 308 tracks for p50-expressing embryos, n = 593 tracks for Kif5C-expressing embryos, n = 441 tracks Kif5C T94N-expressing embryos, n = 373 tracks for Rab11a-expressing embryos, and n = 152 tracks for Rab11a S25N-expressing embryos. Instantaneous velocities in the anterograde (B) as well as retrograde (C) direction are reduced when vesicle formation or motor transport is impaired. For full statistical testing, see Table S3 and Table S4. Anterograde: n = 2,728 values for drug control, n = 2,978 values for Brefeldin A, n = 1,451 values for Dynasore, n = 2,059 values for injection control, n = 1,044 values for p50-expressing embryos, n = 2,808 values for Kif5C-expressing embryos, n = 2,055 values Kif5C T94N-expressing embryos, n = 1,815 values for Rab11a-expressing embryos, and n = 837 values for Rab11a S25N-expressing embryos. Retrograde: n = 2,687 values for drug control, n = 3,034 values for Brefeldin A, n = 1,574 values for Dynasore, n = 2,127 values for injection control, n = 1,117 values for p50-expressing embryos, n = 2,791 values for Kif5C-expressing embryos, n = 2,132 values Kif5C T94N-expressing embryos, n = 1,740 values for Rab11a-expressing embryos, and n = 801 values for Rab11a S25N-expressing embryos. (D) Plotting maximal and mean velocities for each of the 416 particle tracks in control THNs illustrates that the majority of Cadherin-2 particles move only at low mean and maximal velocities, with some particles exhibiting higher mobility in both directions. (E) Converting the mean-maximal velocities plots into data density plots reveals that the fraction of slow-moving particles increases when vesicle formation is impaired by drugs. Color code is indicated on the right, where dark red indicates high data density. For comparison of the spread in the datasets, the area of the 0.8 density is outline in gray for each condition, and the area covered by this density level is indicated in the top left corner. The number of particle tracks for each condition is listed in the bottom left corner. Vertical gray lines separate areas without data content. (F) Similar results are obtained when vesicle formation or motor transport of Cadherin-2 particles are genetically targeted. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. All data shown in A–F represent n = 13 embryos for drug control, n = 19 embryos for Brefeldin A, n = 8 embryos for Dynasore, n = 13 embryos for injection control, n = 14 p50-expressing embryos, n = 11 Kif5C-expressing embryos, n = 16 Kif5C T94N-expressing embryos, n = 16 Rab11a-expressing embryos, and n = 7 Rab11a S25N-expressing embryos.

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